Generation of plants with altered protein, fiber, or oil content

ABSTRACT

The present invention is directed to plants that display an improved oil quantity phenotype or an improved meal quality phenotype due to altered expression of an IMQ nucleic acid. The invention is further directed to methods of generating plants with an improved oil quantity phenotype or improved meal quality phenotype.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of co-pending U.S. applicationSer. No. 13/727,425, filed Dec. 26, 2012; which is a divisional of U.S.application Ser. No. 13/401,682, filed Feb. 21, 2012, now U.S. Pat. No.8,367,892, issued Feb. 5, 2013; which is a divisional of U.S. patentapplication Ser. No. 13/078,843, filed Apr. 1, 2011, now U.S. Pat. No.8,163,978, issued Apr. 24, 2012; which is a divisional of U.S. patentapplication Ser. No. 12/814,412, filed Jun. 11, 2010, now U.S. Pat. No.7,943,817, issued May 17, 2011; which is a divisional of U.S. patentapplication Ser. No. 11/940,284, filed Nov. 14, 2007, now U.S. Pat. No.7,763,771, issued Jul. 27, 2010; which claims the benefit of U.S.Provisional Application No. 60/866,060, filed Nov. 15, 2006; all ofwhich are incorporated herein by reference in their entirety.

FIELD OF THE DISCLOSURE

The present disclosure is related to transgenic plants with altered oil,protein, and/or fiber content, as well as methods of making plantshaving altered oil, protein, and/or fiber content and producing oil fromsuch plants.

BACKGROUND

The ability to manipulate the composition of crop seeds, particularlythe content and composition of seed oil and protein, as well as theavailable metabolizable energy (“AME”) in the seed meal in livestock,has important applications in the agricultural industries, relating bothto processed food oils and to animal feeds. Seeds of agricultural cropscontain a variety of valuable constituents, including oil, protein andstarch. Industrial processing can separate some or all of theseconstituents for individual sale in specific applications. For instance,nearly 60% of the U.S. soybean crop is crushed by the soy processingindustry. Soy processing yields purified oil, which is sold at highvalue, while the remaining seed meal is sold for livestock feed (U.S.Soybean Board, 2001 Soy Stats). Canola seed is also crushed to produceoil and the co-product canola meal (Canola Council of Canada). Canolameal contains a high percentage of protein and a good balance of aminoacids but because it has a high fiber and phytate content, it is notreadily digested by livestock (Slominski et al., 1999 Proceedings of the10^(th) International Rapeseed Congress, Canberra, Australia) and has alower value than soybean meal.

Over 55% of the corn produced in the U.S. is used as animal feed (IowaCorn Growers Association). The value of the corn is directly related toits ability to be digested by livestock. Thus, it is desirable tomaximize both oil content of seeds and the AME of meal. For processedoilseeds such as soy and canola, increasing the absolute oil content ofthe seed will increase the value of such grains, while increasing theAME of meal will increase its value. For processed corn, either anincrease or a decrease in oil content may be desired, depending on howthe other major constituents are to be used. Decreasing oil may improvethe quality of isolated starch by reducing undesired flavors associatedwith oil oxidation. Alternatively, when the starch is used for ethanolproduction, where flavor is unimportant, increasing oil content mayincrease overall value.

In many feed grains, such as corn and wheat, it is desirable to increaseseed oil content, because oil has higher energy content than other seedconstituents such as carbohydrate. Oilseed processing, like most grainprocessing businesses, is a capital-intensive business; thus smallshifts in the distribution of products from the low valued components tothe high value oil component can have substantial economic impacts forgrain processors. In addition, increasing the AME of meal by adjustingseed protein and fiber content and composition, without decreasing seedoil content, can increase the value of animal feed.

Biotechnological manipulation of oils has been shown to providecompositional alteration and improvement of oil yield. Compositionalalterations include high oleic acid soybean and corn oil (U.S. Pat. Nos.6,229,033 and 6,248,939), and laurate-containing seeds (U.S. Pat. No.5,639,790), among others. Work in compositional alteration haspredominantly focused on processed oilseeds, but has been readilyextendable to non-oilseed crops, including corn. While there isconsiderable interest in increasing oil content, the only currentlypracticed biotechnology in this area is High-Oil Corn (HOC) technology(DuPont, U.S. Pat. No. 5,704,160). HOC employs high oil pollinatorsdeveloped by classical selection breeding along with elite(male-sterile) hybrid females in a production system referred to asTopCross. The TopCross High Oil system raises harvested grain oilcontent in maize from about 3.5% to about 7%, improving the energycontent of the grain.

While it has been fruitful, the HOC production system has inherentlimitations. First, the system of having a low percentage of pollinatorsresponsible for an entire field's seed set contains inherent risks,particularly in drought years. Second, oil content in current HOC fieldshas plateaued at about 9% oil. Finally, high-oil corn is not primarily abiochemical change, but rather an anatomical mutant (increased embryosize) that has the indirect result of increasing oil content. For thesereasons, an alternative high oil strategy, particularly one that derivesfrom an altered biochemical output, would be especially valuable.

Manipulation of seed composition has identified several components thatimprove the nutritive quality, digestibility, and AME in seed meal.Increasing the lysine content in canola and soybean (Falco et al., 1995Bio/Technology 13:577-582) increases the availability of this essentialamino acid and decreases the need for nutritional supplements. Soybeanvarieties with increased seed protein were shown to contain considerablymore metabolizable energy than conventional varieties (Edwards et al.,1999, Poultry Sci. 79:525-527). Decreasing the phytate content of cornseed has been shown to increase the bioavailability of amino acids inanimal feeds (Douglas et al., 2000, Poultry Sci. 79:1586-1591) anddecreasing oligosaccharide content in soybean meal increases themetabolizable energy in the meal (Parsons et al., 2000, Poultry Sci.79:1127-1131).

Soybean and canola are the most obvious target crops for the processedoil and seed meal markets since both crops are crushed for oil and theremaining meal sold for animal feed. A large body of commercial work(e.g., U.S. Pat. No. 5,952,544; PCT Application No. WO9411516)demonstrates that Arabidopsis is an excellent model for oil metabolismin these crops. Biochemical screens of seed oil composition haveidentified Arabidopsis genes for many critical biosynthetic enzymes andhave led to identification of agronomically important gene orthologs.For instance, screens using chemically mutagenized populations haveidentified lipid mutants whose seeds display altered fatty acidcomposition (Lemieux et al., 1990, Theor. Appl. Genet. 80, 234-240;James and Dooner, 1990, Theor. Appl. Genet. 80, 241-245). T-DNAmutagenesis screens (Feldmann et al., 1989, Science 243: 1351-1354) thatdetected altered fatty acid composition identified the omega 3desaturase (FADS) and delta-12 desaturase (FAD2) genes (U.S. Pat. No.5,952,544; Yadav et al., 1993, Plant Physiol. 103, 467-476; Okuley etal., 1994, Plant Cell 6(1):147-158). A screen which focused on oilcontent rather than oil quality, analyzed chemically-induced mutants forwrinkled seeds or altered seed density, from which altered seed oilcontent was inferred (Focks and Benning, 1998, Plant Physiol.118:91-101).

Another screen, designed to identify enzymes involved in production ofvery long chain fatty acids, identified a mutation in the gene encodinga diacylglycerol acyltransferase (DGAT) as being responsible for reducedtriacyl glycerol accumulation in seeds (Katavic V et al., 1995, PlantPhysiol. 108(1):399-409). It was further shown that seed-specificover-expression of the DGAT cDNA was associated with increased seed oilcontent (Jako et al., 2001, Plant Physiol. 126(2):861-74). Arabidopsisis also a model for understanding the accumulation of seed componentsthat affect meal quality. For example, Arabidopsis contains albumin andglobulin seed storage proteins found in many dicotyledonous plantsincluding canola and soybean (Shewry 1995, Plant Cell 7:945-956). Thebiochemical pathways for synthesizing components of fiber, such ascellulose and lignin, are conserved within the vascular plants, andmutants of Arabidopsis affecting these components have been isolated(reviewed in Chapel and Carpita 1998, Current Opinion in Plant Biology1:179-185).

Activation tagging in plants refers to a method of generating randommutations by insertion of a heterologous nucleic acid constructcomprising regulatory sequences (e.g., an enhancer) into a plant genome.The regulatory sequences can act to enhance transcription of one or morenative plant genes; accordingly, activation tagging is a fruitful methodfor generating gain-of-function, generally dominant mutants (see, e.g.,Hayashi et al., 1992, Science 258: 1350-1353; Weigel D et al., 2000,Plant Physiology, 122:1003-1013). The inserted construct provides amolecular tag for rapid identification of the native plant whosemis-expression causes the mutant phenotype. Activation tagging may alsocause loss-of-function phenotypes. The insertion may result indisruption of a native plant gene, in which case the phenotype isgenerally recessive.

Activation tagging has been used in various species, including tobaccoand Arabidopsis, to identify many different kinds of mutant phenotypesand the genes associated with these phenotypes (Wilson et al., 1996,Plant Cell 8: 659-671; Schaffer et al., 1998, Cell 93: 1219-1229;Fridborg et al., 1999, Plant Cell 11: 1019-1032; Kardailsky et al.,1999, Science 286: 1962-1965; and Christensen S et al., 1998, 9^(th)International Conference on Arabidopsis Research, Univ. ofWisconsin-Madison, June 24-28, Abstract 165).

SUMMARY

Provided herein are transgenic plants having an Improved Seed Qualityphenotype. Transgenic plants with an Improved Seed Quality phenotype mayinclude an improved oil quantity and/or an improved meal quality.Transgenic plants with improved meal quality have an Improved MealQuality (IMQ) phenotype and transgenic plants with improved oil quantityhave an Improved Oil Quantity (IOQ) phenotype. The IMQ phenotype in atransgenic plant may include altered protein and/or fiber content in anypart of the transgenic plant, for example in the seeds. The IOQphenotype in a transgenic plant may include altered oil content in anypart of the transgenic plant, for example in the seeds. In particularembodiments, a transgenic plant may include an IOQ phenotype and/or anIMQ phenotype. In some embodiments of a transgenic plant, the IMQphenotype may be an increase in protein content in the seed and/or adecrease in the fiber content of the seed. In other embodiments of atransgenic plant, the IOQ phenotype is an increase in the oil content ofthe seed (a high oil phenotype). Also provided is seed meal derived fromthe seeds of transgenic plants, wherein the seeds have altered proteincontent and/or altered fiber content. Further provided is oil derivedfrom the seeds of transgenic plants, wherein the seeds have altered oilcontent. Any of these changes can lead to an increase in the AME fromthe seed or seed meal from transgenic plants, relative to control,non-transgenic, or wild-type plants. Also provided herein is meal, feed,or food produced from any part of the transgenic plant with an IMQphenotype and/or IOQ phenotype.

In certain embodiments, the disclosed transgenic plants comprise atransformation vector comprising an IMQ nucleotide sequence that encodesor is complementary to a sequence that encodes an “IMQ” polypeptide. Inparticular embodiments, expression of an IMQ polypeptide in a transgenicplant causes an altered oil content, an altered protein content, and/oran altered fiber content in the transgenic plant. In preferredembodiments, the transgenic plant is selected from the group consistingof plants of the Brassica species, including canola and rapeseed, soy,corn, sunflower, cotton, cocoa, safflower, oil palm, coconut palm, flax,castor, peanut, wheat, oat and rice. Also provided is a method ofproducing oil or seed meal, comprising growing the transgenic plant andrecovering oil and/or seed meal from said plant. The disclosure furtherprovides feed, meal, grain, or seed comprising a nucleic acid sequencethat encodes an IMQ polypeptide. The disclosure also provides feed,meal, grain, or seed comprising the IMQ polypeptide, or an orthologthereof.

Examples of the disclosed transgenic plant are produced by a method thatcomprises introducing into progenitor cells of the plant a planttransformation vector comprising an IMQ nucleotide sequence thatencodes, or is complementary to a sequence that encodes, an IMQpolypeptide, and growing the transformed progenitor cells to produce atransgenic plant, wherein the IMQ polynucleotide sequence is expressed,causing an IOQ phenotype and/or and IMQ phenotype in the transgenicplant. In some specific, non-limiting examples, the method producestransgenic plants wherein expression of the IMQ polypeptide causes ahigh (increased) oil, high (increased) protein, and/or low (decreased)fiber phenotype in the transgenic plant, relative to control,non-transgenic, or wild-type plants.

Additional methods are disclosed herein of generating a plant having anIMQ and/or an IOQ phenotype, wherein a plant is identified that has anallele in its IMQ nucleic acid sequence that results in an IMQ phenotypeand/or an IOQ phenotype, compared to plants lacking the allele. Theplant can generate progeny, wherein the progeny inherit the allele andhave an IMQ phenotype and/or an IOQ phenotype. In some embodiments ofthe method, the method employs candidate gene/QTL methodology or TILLINGmethodology.

Also provided herein is a transgenic plant cell having an IMQ phenotypeand/or an IOQ phenotype. The transgenic plant cell comprises atransformation vector comprising an IMQ nucleotide sequence that encodesor is complementary to a sequence that encodes an IMQ polypeptide. Inpreferred embodiments, the transgenic plant cell is selected from thegroup consisting of plants of the Brassica species, including canola andrapeseed, soy, corn, sunflower, cotton, cocoa, safflower, oil palm,coconut palm, flax, castor, peanut, wheat, oat and rice. In otherembodiments, the plant cell is a seed, pollen, propagule, or embryocell. The disclosure also provides plant cells from a plant that is thedirect progeny or the indirect progeny of a plant grown from saidprogenitor cells.

INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED AS AN ASCII TEXT FILE

The Sequence Listing is submitted as an ASCII text file in the form ofthe file named Sequence_Listing.txt, which was created on Sep. 18, 2015,and is 410,962 bytes, which is incorporated by reference herein it isentirety.

DETAILED DESCRIPTION Terms

Unless otherwise indicated, all technical and scientific terms usedherein have the same meaning as they would to one skilled in the art ofthe present disclosure. Practitioners are particularly directed toSambrook et al. (Molecular Cloning: A Laboratory Manual (SecondEdition), Cold Spring Harbor Press, Plainview, N. Y., 1989) and AusubelF M et al. (Current Protocols in Molecular Biology, John Wiley & Sons,New York, N. Y., 1993) for definitions and terms of the art. It is to beunderstood that this disclosure is not limited to the particularmethodology, protocols, and reagents described, as these may vary.

As used herein, the term “IMQ phenotype” refers to plants, or any partof a plant (for example, seeds, or meal produced from seeds), with analtered protein and/or fiber content (phenotype). As provided herein,altered protein and/or fiber content includes either an increased ordecreased level of protein and/or fiber content in plants, seeds or seedmeal. Any combination of these changes can lead to an IMQ phenotype. Forexample, in one specific non-limiting example, an IMQ phenotype canrefer to increased protein and decreased fiber content. In anotherspecific non-limiting example, an IMQ phenotype can refer to unchangedprotein and decreased fiber content. In yet another specificnon-limiting example, an IMQ phenotype can refer to increased proteinand unchanged fiber content. It is also provided that any combination ofthese changes can lead to an increase in the AME (availablemetabolizable energy) from the seed or meal generated from the seed. AnIMQ phenotype also includes an improved seed quality (ISQ) phenotype oran improved seed meal quality phenotype.

As used herein, the term “IOQ phenotype” refers to plants, or any partof a plant (for example, seeds), with an altered oil content(phenotype). As provided herein, altered oil content includes anincreased, for example a high, oil content in plants or seeds. In someembodiments, a transgenic plant can express both an IOQ phenotype and anIMQ phenotype. In specific, non-limiting examples, a transgenic planthaving a combination of an IOQ phenotype and an IMQ phenotype can leadto an increase in the AME (available metabolizable energy) from the seedor meal generated from the seed. An IOQ phenotype also includes animproved seed quality (ISQ) phenotype.

As used herein, the term “available metabolizable energy” (AME) refersto the amount of energy in the feed that is able to be extracted bydigestion in an animal and is correlated with the amount of digestibleprotein and oil available in animal meal. AME is determined byestimating the amount of energy in the feed prior to feeding andmeasuring the amount of energy in the excreta of the animal followingconsumption of the feed. In one specific, non-limiting example, atransgenic plant with an increase in AME includes transgenic plants withaltered seed protein and/or fiber content and without a decrease in seedoil content (seed oil content remains unchanged or is increased),resulting in an increase in the value of animal feed derived from theseed.

As used herein, the term “content” refers to the type and relativeamount of, for instance, a seed or seed meal component.

As used herein, the term “fiber” refers to non-digestible components ofthe plant seed including cellular components such as cellulose,hemicellulose, pectin, lignin, and phenolics.

As used herein, the term “meal” refers to seed components remainingfollowing the extraction of oil from the seed. Examples of components ofmeal include protein and fiber.

As used herein, the term “vector” refers to a nucleic acid constructdesigned for transfer between different host cells. An “expressionvector” refers to a vector that has the ability to incorporate andexpress heterologous DNA fragments in a foreign cell. Many prokaryoticand eukaryotic expression vectors are commercially available. Selectionof appropriate expression vectors is within the knowledge of thosehaving skill in the art.

A “heterologous” nucleic acid construct or sequence has a portion of thesequence that is not native to the plant cell in which it is expressed.Heterologous, with respect to a control sequence refers to a controlsequence (i.e. promoter or enhancer) that does not function in nature toregulate the same gene the expression of which it is currentlyregulating. Generally, heterologous nucleic acid sequences are notendogenous to the cell or part of the genome in which they are present,and have been added to the cell by infection, transfection,microinjection, electroporation, or the like. A “heterologous” nucleicacid construct may contain a control sequence/DNA coding sequencecombination that is the same as, or different from, a controlsequence/DNA coding sequence combination found in the native plant.Specific, non-limiting examples of a heterologous nucleic acid sequenceinclude an IMQ nucleic acid sequence, or a fragment, derivative(variant), or ortholog thereof.

As used herein, the term “gene” means the segment of DNA involved inproducing a polypeptide chain, which may or may not include regionspreceding and following the coding region, e.g. 5′ untranslated (5′ UTR)or “leader” sequences and 3′ UTR or “trailer” sequences, as well asintervening sequences (introns) between individual coding segments(exons) and non-transcribed regulatory sequences.

As used herein, “recombinant” includes reference to a cell or vector,that has been modified by the introduction of a heterologous nucleicacid sequence or that the cell is derived from a cell so modified. Thus,for example, recombinant cells express genes that are not found inidentical form within the native (non-recombinant) form of the cell orexpress native genes that are otherwise abnormally expressed, underexpressed, or not expressed at all as a result of deliberate humanintervention.

As used herein, the term “gene expression” refers to the process bywhich a polypeptide is produced based on the nucleic acid sequence of agene. The process includes both transcription and translation;accordingly, “expression” may refer to either a polynucleotide orpolypeptide sequence, or both. Sometimes, expression of a polynucleotidesequence will not lead to protein translation. “Over-expression” refersto increased expression of a polynucleotide and/or polypeptide sequencerelative to its expression in a wild-type (or other reference [e.g.,non-transgenic]) plant and may relate to a naturally-occurring ornon-naturally occurring sequence. “Ectopic expression” refers toexpression at a time, place, and/or increased level that does notnaturally occur in the non-altered or wild-type plant.“Under-expression” refers to decreased expression of a polynucleotideand/or polypeptide sequence, generally of an endogenous gene, relativeto its expression in a wild-type plant. The terms “mis-expression” and“altered expression” encompass over-expression, under-expression, andectopic expression.

The term “introduced” in the context of inserting a nucleic acidsequence into a cell, includes “transfection,” “transformation,” and“transduction” and includes reference to the incorporation of a nucleicacid sequence into a eukaryotic or prokaryotic cell where the nucleicacid sequence may be incorporated into the genome of the cell (forexample, chromosome, plasmid, plastid, or mitochondrial DNA), convertedinto an autonomous replicon, or transiently expressed (for example,transfected mRNA).

As used herein, a “plant cell” refers to any cell derived from a plant,including cells from undifferentiated tissue (e.g., callus), as well asfrom plant seeds, pollen, propagules, and embryos.

As used herein, the terms “native” and “wild-type” relative to a givenplant trait or phenotype refers to the form in which that trait orphenotype is found in the same variety of plant in nature. In oneembodiment, a wild-type plant is also a control plant. In anotherembodiment, a wild-type plant is a non-transgenic plant.

As used herein, the term “modified” regarding a plant trait, refers to achange in the phenotype of a transgenic plant (for example, a transgenicplant with any combination of an altered oil content, an altered proteincontent, and/or an altered fiber content) in any part of the transgenicplant, for example the seeds, relative to a similar non-transgenicplant. As used herein, the term “altered” refers to either an increaseor a decrease of a plant trait or phenotype (for example, oil content,protein content, and/or fiber content) in a transgenic plant, relativeto a similar non-transgenic plant. In one specific, non-limitingexample, a transgenic plant with a modified trait includes a plant withan increased oil content, increased protein content, and/or decreasedfiber content relative to a similar non-transgenic plant. In anotherspecific, non-limiting example, a transgenic plant with a modified traitincludes unchanged oil content, increased protein content, and/ordecreased fiber content relative to a similar non-transgenic plant. Inyet another specific, non-limiting example, a transgenic plant with amodified trait includes an increased oil content, increased proteincontent, and/or unchanged fiber content relative to a similarnon-transgenic plant. Specific, non-limiting examples of a change inphenotype include an IMQ phenotype or an IOQ phenotype.

An “interesting phenotype (trait)” with reference to a transgenic plantrefers to an observable or measurable phenotype demonstrated by a T1and/or subsequent generation plant, which is not displayed by thecorresponding non-transgenic plant (i.e., a genotypically similar plantthat has been raised or assayed under similar conditions). Aninteresting phenotype may represent an improvement in the plant (forexample, increased oil content, increased protein content, and/ordecreased fiber content in seeds of the plant) or may provide a means toproduce improvements in other plants. An “improvement” is a feature thatmay enhance the utility of a plant species or variety by providing theplant with a unique and/or novel phenotype or quality. Such transgenicplants may have an improved phenotype, such as an IMQ phenotype or anIOQ phenotype.

The phrase “altered oil content phenotype” refers to a measurablephenotype of a genetically modified (transgenic) plant, where the plantdisplays a statistically significant increase or decrease in overall oilcontent (i.e., the percentage of seed mass that is oil), as compared tothe similar, but non-modified (non-transgenic) plant. A high oilphenotype refers to an increase in overall oil content. The phrase“altered protein content phenotype” refers to measurable phenotype of agenetically modified plant, where the plant displays a statisticallysignificant increase or decrease in overall protein content (i.e., thepercentage of seed mass that is protein), as compared to the similar,but non-modified plant. A high protein phenotype refers to an increasein overall protein content. The phrase “altered fiber content phenotype”refers to measurable phenotype of a genetically modified plant, wherethe plant displays a statistically significant increase or decrease inoverall fiber content (i.e., the percentage of seed mass that is fiber),as compared to the similar, but non-modified plant. A low fiberphenotype refers to decrease in overall fiber content.

As used herein, a “mutant” polynucleotide sequence or gene differs fromthe corresponding wild-type polynucleotide sequence or gene either interms of sequence or expression, where the difference contributes to amodified or altered plant phenotype or trait. Relative to a plant orplant line, the term “mutant” refers to a plant or plant line which hasa modified or altered plant phenotype or trait, where the modified oraltered phenotype or trait is associated with the modified or alteredexpression of a wild-type polynucleotide sequence or gene.

As used herein, the term “T1” refers to the generation of plants fromthe seed of T0 plants. The T1 generation is the first set of transformedplants that can be selected by application of a selection agent, e.g.,an antibiotic or herbicide, for which the transgenic plant contains thecorresponding resistance gene. The term “T2” refers to the generation ofplants by self-fertilization of the flowers of T1 plants, previouslyselected as being transgenic. T3 plants are generated from T2 plants,etc. As used herein, the “direct progeny” of a given plant derives fromthe seed (or, sometimes, other tissue) of that plant and is in theimmediately subsequent generation; for instance, for a given lineage, aT2 plant is the direct progeny of a T1 plant. The “indirect progeny” ofa given plant derives from the seed (or other tissue) of the directprogeny of that plant, or from the seed (or other tissue) of subsequentgenerations in that lineage; for instance, a T3 plant is the indirectprogeny of a T1 plant.

As used herein, the term “plant part” includes any plant organ ortissue, including, without limitation, seeds, embryos, meristematicregions, callus tissue, leaves, roots, shoots, gametophytes,sporophytes, pollen, and microspores. Plant cells can be obtained fromany plant organ or tissue and cultures prepared therefrom. Providedherein is a transgenic plant cell having an IMQ phenotype and/or an IOQphenotype. The transgenic plant cell comprises a transformation vectorcomprising an IMQ nucleotide sequence that encodes or is complementaryto a sequence that encodes an IMQ polypeptide. In preferred embodiments,the transgenic plant cell is selected from the group consisting ofplants of the Brassica species, including canola and rapeseed, soy,corn, sunflower, cotton, cocoa, safflower, oil palm, coconut palm, flax,castor, peanut, wheat, oat and rice. In other embodiments, the plantcell is a seed, pollen, propagule, or embryo cell. The disclosure alsoprovides plant cells from a plant that is the direct progeny or theindirect progeny of a plant grown from said progenitor cells. The classof plants which can be used in the methods of the present invention isgenerally as broad as the class of higher plants amenable totransformation techniques, including both monocotyledonous anddicotyledonous plants.

As used herein, “transgenic plant” includes a plant that compriseswithin its genome a heterologous polynucleotide. The heterologouspolynucleotide can be either stably integrated into the genome, or canbe extra-chromosomal. Preferably, the polynucleotide of the presentinvention is stably integrated into the genome such that thepolynucleotide is passed on to successive generations. A plant cell,tissue, organ, or plant into which the heterologous polynucleotides havebeen introduced is considered “transformed,” “transfected,” or“transgenic.” Direct and indirect progeny of transformed plants or plantcells that also contain the heterologous polynucleotide are alsoconsidered transgenic.

Disclosed herein are transgenic plants having an Improved Seed Qualityphenotype. Transgenic plants with an Improved Seed Quality phenotype mayinclude an improved oil quantity and/or an improved meal quality.Transgenic plants with improved meal quality have an IMQ phenotype andtransgenic plants with improved oil quantity have an IOQ phenotype. TheIMQ phenotype in a transgenic plant may include altered protein and/orfiber content in any part of the transgenic plant, for example in theseeds. The IOQ phenotype in a transgenic plant may include altered oilcontent in any part of the transgenic plant, for example in the seeds.In particular embodiments, a transgenic plant may include an IOQphenotype and/or an IMQ phenotype. In some embodiments of a transgenicplant, the IMQ phenotype may be an increase in protein content in theseed and/or a decrease in the fiber content of the seed. In otherembodiments of a transgenic plant, the IOQ phenotype is an increase inthe oil content of the seed (a high oil phenotype). Also provided isseed meal derived from the seeds of transgenic plants, wherein the seedshave altered protein content and/or altered fiber content. Furtherprovided is oil derived from the seeds of transgenic plants, wherein theseeds have altered oil content. Any of these changes can lead to anincrease in the AME from the seed or seed meal from transgenic plants,relative to control, non-transgenic, or wild-type plants. Also providedherein is meal, feed, or food produced from any part of the transgenicplant with an IMQ phenotype and/or IOQ phenotype.

In certain embodiments, the disclosed transgenic plants comprise atransformation vector comprising an IMQ nucleotide sequence that encodesor is complementary to a sequence that encodes an “IMQ” polypeptide. Inparticular embodiments, expression of an IMQ polypeptide in a transgenicplant causes an altered oil content, an altered protein content, and/oran altered fiber content in the transgenic plant. In preferredembodiments, the transgenic plant is selected from the group consistingof plants of the Brassica species, including canola and rapeseed, soy,corn, sunflower, cotton, cocoa, safflower, oil palm, coconut palm, flax,castor, peanut, wheat, oat and rice. Also provided is a method ofproducing oil or seed meal, comprising growing the transgenic plant andrecovering oil and/or seed meal from said plant. The disclosure furtherprovides feed, meal, grain, or seed comprising a nucleic acid sequencethat encodes an IMQ polypeptide. The disclosure also provides feed,meal, grain, or seed comprising the IMQ polypeptide, or an orthologthereof.

Various methods for the introduction of a desired polynucleotidesequence encoding the desired protein into plant cells are available andknown to those of skill in the art and include, but are not limited to:(1) physical methods such as microinjection, electroporation, andmicroprojectile mediated delivery (biolistics or gene gun technology);(2) virus mediated delivery methods; and (3) Agrobacterium-mediatedtransformation methods (see, for example, WO 2007/053482 and WO2005/107437, which are incorporated herein by reference in theirentirety).

The most commonly used methods for transformation of plant cells are theAgrobacterium-mediated DNA transfer process and the biolistics ormicroprojectile bombardment mediated process (i.e., the gene gun).Typically, nuclear transformation is desired but where it is desirableto specifically transform plastids, such as chloroplasts or amyloplasts,plant plastids may be transformed utilizing a microprojectile-mediateddelivery of the desired polynucleotide.

Agrobacterium-mediated transformation is achieved through the use of agenetically engineered soil bacterium belonging to the genusAgrobacterium. A number of wild-type and disarmed strains ofAgrobacterium tumefaciens and Agrobacterium rhizogenes harboring Ti orRi plasmids can be used for gene transfer into plants. Gene transfer isdone via the transfer of a specific DNA known as “T-DNA” that can begenetically engineered to carry any desired piece of DNA into many plantspecies.

Agrobacterium-mediated genetic transformation of plants involves severalsteps. The first step, in which the virulent Agrobacterium and plantcells are first brought into contact with each other, is generallycalled “inoculation.” Following the inoculation, the Agrobacterium andplant cells/tissues are permitted to be grown together for a period ofseveral hours to several days or more under conditions suitable forgrowth and T-DNA transfer. This step is termed “co-culture.” Followingco-culture and T-DNA delivery, the plant cells are treated withbactericidal or bacteriostatic agents to kill the Agrobacteriumremaining in contact with the explant and/or in the vessel containingthe explant. If this is done in the absence of any selective agents topromote preferential growth of transgenic versus non-transgenic plantcells, then this is typically referred to as the “delay” step. If donein the presence of selective pressure favoring transgenic plant cells,then it is referred to as a “selection” step. When a “delay” is used, itis typically followed by one or more “selection” steps.

With respect to microprojectile bombardment (U.S. Pat. No. 5,550,318;U.S. Pat. No. 5,538,880, U.S. Pat. No. 5,610,042; and PCT Publication WO95/06128; each of which is specifically incorporated herein by referencein its entirety), particles are coated with nucleic acids and deliveredinto cells by a propelling force. Exemplary particles include thosecomprised of tungsten, platinum, and preferably, gold.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System(BioRad, Hercules, Calif.), which can be used to propel particles coatedwith DNA or cells through a screen, such as a stainless steel or Nytexscreen, onto a filter surface covered with monocot plant cells culturedin suspension.

Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species. Examples of species thathave been transformed by microprojectile bombardment include monocotspecies such as maize (PCT Publication No. WO 95/06128), barley, wheat(U.S. Pat. No. 5,563,055, incorporated herein by reference in itsentirety), rice, oat, rye, sugarcane, and sorghum, as well as a numberof dicots including tobacco, soybean (U.S. Pat. No. 5,322,783,incorporated herein by reference in its entirety), sunflower, peanut,cotton, tomato, and legumes in general (U.S. Pat. No. 5,563,055,incorporated herein by reference in its entirety).

To select or score for transformed plant cells regardless oftransformation methodology, the DNA introduced into the cell contains agene that functions in a regenerable plant tissue to produce a compoundthat confers upon the plant tissue resistance to an otherwise toxiccompound. Genes of interest for use as a selectable, screenable, orscorable marker would include but are not limited to GUS, greenfluorescent protein (GFP), luciferase (LUX), antibiotic or herbicidetolerance genes. Examples of antibiotic resistance genes include thepenicillins, kanamycin (and neomycin, G418, bleomycin), methotrexate(and trimethoprim), chloramphenicol, and tetracycline. Polynucleotidemolecules encoding proteins involved in herbicide tolerance are known inthe art, and include, but are not limited to a polynucleotide moleculeencoding 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) describedin U.S. Pat. No. 5,627,061, U.S. Pat. No. 5,633,435, and U.S. Pat. No.6,040,497 and aroA described in U.S. Pat. No. 5,094,945 for glyphosatetolerance; a polynucleotide molecule encoding bromoxynil nitrilase (Bxn)described in U.S. Pat. No. 4,810,648 for Bromoxynil tolerance; apolynucleotide molecule encoding phytoene desaturase (crtl) described inMisawa et al., (Plant J. 4:833-840, 1993) and Misawa et al., (Plant J.6:481-489, 1994) for norflurazon tolerance; a polynucleotide moleculeencoding acetohydroxyacid synthase (AHAS, also known as ALS) describedin Sathasiivan et al. (Nucl. Acids Res. 18:2188-2193, 1990) fortolerance to sulfonylurea herbicides; and the bar gene described inDeBlock, et al., (EMBO J. 6:2513-2519, 1987) for glufosinate andbialaphos tolerance.

The regeneration, development, and cultivation of plants from varioustransformed explants are well documented in the art. This regenerationand growth process typically includes the steps of selecting transformedcells and culturing those individualized cells through the usual stagesof embryonic development through the rooted plantlet stage. Transgenicembryos and seeds are similarly regenerated. The resulting transgenicrooted shoots are thereafter planted in an appropriate plant growthmedium such as soil. Cells that survive the exposure to the selectiveagent, or cells that have been scored positive in a screening assay, maybe cultured in media that supports regeneration of plants. Developingplantlets are transferred to soil less plant growth mix, and hardenedoff, prior to transfer to a greenhouse or growth chamber for maturation.

The present invention can be used with any transformable cell or tissue.By transformable as used herein is meant a cell or tissue that iscapable of further propagation to give rise to a plant. Those of skillin the art recognize that a number of plant cells or tissues aretransformable in which after insertion of exogenous DNA and appropriateculture conditions the plant cells or tissues can form into adifferentiated plant. Tissue suitable for these purposes can include butis not limited to immature embryos, scutellar tissue, suspension cellcultures, immature inflorescence, shoot meristem, nodal explants, callustissue, hypocotyl tissue, cotyledons, roots, and leaves.

Any suitable plant culture medium can be used. Examples of suitablemedia would include but are not limited to MS-based media (Murashige andSkoog, Physiol. Plant, 15:473-497, 1962) or N6-based media (Chu et al.,Scientia Sinica 18:659, 1975) supplemented with additional plant growthregulators including but not limited to auxins, cytokinins, ABA, andgibberellins. Those of skill in the art are familiar with the variety oftissue culture media, which when supplemented appropriately, supportplant tissue growth and development and are suitable for planttransformation and regeneration. These tissue culture media can eitherbe purchased as a commercial preparation, or custom prepared andmodified. Those of skill in the art are aware that media and mediasupplements such as nutrients and growth regulators for use intransformation and regeneration and other culture conditions such aslight intensity during incubation, pH, and incubation temperatures thatcan be optimized for the particular variety of interest.

One of ordinary skill will appreciate that, after an expression cassetteis stably incorporated in transgenic plants and confirmed to beoperable, it can be introduced into other plants by sexual crossing. Anyof a number of standard breeding techniques can be used, depending uponthe species to be crossed.

Identification of Plants with an Improved Oil Quantity Phenotype and/orImproved Meal Quality Phenotype

An Arabidopsis activation tagging screen (ACTTAG) was used to identifythe association between 1) ACTTAG plant lines with an altered protein,fiber and/or oil content (phenotype, for example, see columns 4, 5 and6, respectively, of Table 1, below) and 2) the nucleic acid sequencesidentified in column 3 of Tables 2 and 3, wherein each nucleic acidsequence is provided with a gene alias or an IMQ designation (IMQ#; seecolumn 1 in Tables 1, 2, and 3). Briefly, and as further described inthe Examples, a large number of Arabidopsis plants were mutated with thepSKI015 vector, which comprises a T-DNA from the Ti plasmid ofAgrobacterium tumefaciens, a viral enhancer element, and a selectablemarker gene (Weigel et al., 2000, Plant Physiology, 122:1003-1013). Whenthe T-DNA inserts into the genome of transformed plants, the enhancerelement can cause up-regulation of genes in the vicinity, generallywithin about nine kilobases (kb) of the enhancers. T1 plants wereexposed to the selective agent in order to specifically recovertransformed plants that expressed the selectable marker and thereforeharbored T-DNA insertions. T1 plants were allowed to grow to maturity,self-fertilize and produce seed. T2 seed was harvested, labeled andstored. To amplify the seed stocks, about eighteen T2 were sown in soiland, after germination, exposed to the selective agent to recovertransformed T2 plants. T3 seed from these plants was harvested andpooled. Oil, protein and fiber content of the seed were estimated usingNear Infrared Spectroscopy (NIR) as described in the Examples.

Quantitative determination of fatty acid (FA) content (column 7,Table 1) in T2 seeds was performed using the following methods. A sampleof 15 to 20 T2 seeds from each line tested. This sample generallycontained plants with homozygous insertions, no insertions, andhemizygous insertions in a standard 1:1:2 ratios. The seed sample wasmassed on UMT-2 ultra-microbalance (Mettler-Toledo Co., OH) and thentransferred to a glass extraction vial. Lipids were extracted from theseeds and trans-esterified in 500 μl 2.5% H₂SO₄ in MeOH for 3 hours at80° C., following the method of Browse et al. (Biochem J 235:25-31,1986) with modifications. A known amount of heptadecanoic acid wasincluded in the reaction as an internal standard. 750 μl of water and400 μl of hexane were added to each vial, which was then shakenvigorously and allowed to phase separate. Reaction vials were loadeddirectly onto gas chromatography (GC) for analysis and the upper hexanephase was sampled by the autosampler. Gas chromatography with FlameIonization detection was used to separate and quantify the fatty acidmethyl esters. Agilent 6890 Plus GC's were used for separation withAgilent Innowax columns (30 m×0.25 mm ID, 250 μm film thickness). Thecarrier gas was Hydrogen at a constant flow of 2.5 ml/minute. 1 μl ofsample was injected in splitless mode (inlet temperature 220° C., Purgeflow 15 ml/min at 1 minute). The oven was programmed for an initialtemperature of 105° C., initial time 0.5 minutes, followed by a ramp of60° C. per minute to 175° C., a 40° C. /minute ramp to 260° C. with afinal hold time of 2 minutes. Detection was by Flame Ionization(Temperature 275° C., Fuel flow 30.0 ml/min, Oxidizer 400.0 ml/min).Instrument control and data collection and analysis were monitored usingthe Millennium Chromatography Management System (Version 3.2, WatersCorporation, Milford, Mass.). Peaks were initially identified bycomparison with standards. Integration and quantification were performedautomatically, but all analyses were subsequently examined manually toverify correct peak identification and acceptable signal to noise ratiobefore inclusion of the derived results in the study.

The association of an IMQ nucleic acid sequence with an IMQ phenotype oran IOQ phenotype was discovered by analysis of the genomic DNA sequenceflanking the T-DNA insertion in the ACTTAG line identified in column 3of Table 1. An ACTTAG line is a family of plants derived from a singleplant that was transformed with a T-DNA element containing four tandemcopies of the CaMV 35S enhancers. Accordingly, the disclosed IMQ nucleicacid sequences and/or polypeptides may be employed in the development oftransgenic plants having an improved seed quality phenotype, includingan IMQ phenotype and/or an IOQ phenotype. IMQ nucleic acid sequences maybe used in the generation of transgenic plants, such as oilseed crops,that provide improved oil yield from oilseed processing and result in anincrease in the quantity of oil recovered from seeds of the transgenicplant. IMQ nucleic acid sequences may also be used in the generation oftransgenic plants, such as feed grain crops, that provide an IMQphenotype resulting in increased energy for animal feeding, for example,seeds or seed meal with an altered protein and/or fiber content,resulting in an increase in AME. IMQ nucleic acid sequences may furtherbe used to increase the oil content of specialty oil crops, in order toaugment yield and/or recovery of desired unusual fatty acids. Transgenicplants that have been genetically modified to express IMQ polypeptidescan be used in the production of seeds, wherein the transgenic plantsare grown, and oil and seed meal are obtained from plant parts (e.g.seed) using standard methods.

IMQ Nucleic Acids and Polypeptides

The IMQ designation for each of the IMQ nucleic acid sequencesdiscovered in the activation tagging screen described herein are listedin column 1 of Tables 1-3, below. The disclosed IMQ polypeptides arelisted in column 5 of Table 2 and column 4 of Table 3. As used herein,the term “IMQ polypeptide” refers to any polypeptide that when expressedin a plant causes an IMQ phenotype and/or an IOQ phenotype in any partof the plant, for example the seeds. In one embodiment, an IMQpolypeptide refers to a full-length IMQ protein, or a fragment,derivative (variant), or ortholog thereof that is “functionally active,”such that the protein fragment, derivative, or ortholog exhibits one ormore or the functional activities associated with one or more of thedisclosed full-length IMQ polypeptides, for example, the amino acidsequences provided in the GenBank entry referenced in column 5 of Table2, which correspond to the amino acid sequences set forth as SEQ ID NO:2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO:12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ IDNO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40,SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO:50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ IDNO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78,SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO:88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ IDNO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106,SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ IDNO: 116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122, or anortholog thereof. In one preferred embodiment, a functionally active IMQpolypeptide causes an IMQ phenotype and/or an IOQ phenotype in atransgenic plant. In another embodiment, a functionally active IMQpolypeptide causes an altered oil, protein, and/or fiber contentphenotype (for example, an altered seed meal content phenotype) whenmis-expressed in a plant. In other preferred embodiments, mis-expressionof the IMQ polypeptide causes a high oil (such as, increased oil), highprotein (such as, increased protein), and/or low fiber (such as,decreased fiber) phenotype in a plant. In another embodiment,mis-expression of the IMQ polypeptide causes an improved AME of meal. Inyet another embodiment, a functionally active IMQ polypeptide can rescuedefective (including deficient) endogenous IMQ activity when expressedin a plant or in plant cells; the rescuing polypeptide may be from thesame or from a different species as the species with the defectivepolypeptide activity. The disclosure also provides feed, meal, grain,food, or seed comprising the IMQ polypeptide, or a fragment, derivative(variant), or ortholog thereof.

In another embodiment, a functionally active fragment of a full lengthIMQ polypeptide (for example, a functionally active fragment of a nativepolypeptide having the amino acid sequence set forth as SEQ ID NO: 2,SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12,SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO:22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ IDNO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50,SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO:60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ IDNO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88,SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO:98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO:116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122, or a naturallyoccurring ortholog thereof) retains one or more of the biologicalproperties associated with the full-length IMQ polypeptide, such assignaling activity, binding activity, catalytic activity, or cellular orextra-cellular localizing activity. An IMQ fragment preferably comprisesan IMQ domain, such as a C- or N-terminal or catalytic domain, amongothers, and preferably comprises at least 10, preferably at least 20,more preferably at least 25, and most preferably at least 50 contiguousamino acids of an IMQ protein. Functional domains of IMQ genes arelisted in column 8 of Table 2 and can be identified using the PFAMprogram (Bateman A et al., 1999, Nucleic Acids Res. 27:260-262) orINTERPRO (Mulder et al., 2003, Nucleic Acids Res. 31, 315-318) program.Functionally active variants of full-length IMQ polypeptides, orfragments thereof, include polypeptides with amino acid insertions,deletions, or substitutions that retain one of more of the biologicalproperties associated with the full-length IMQ polypeptide. In somecases, variants are generated that change the post-translationalprocessing of an IMQ polypeptide. For instance, variants may havealtered protein transport or protein localization characteristics, oraltered protein half-life, compared to the native polypeptide.

As used herein, the term “IMQ nucleic acid” refers to any polynucleotidethat when expressed in a plant causes an IMQ phenotype and/or an IOQphenotype in any part of the plant, for example the seeds. In oneembodiment, an IMQ polynucleotide encompasses nucleic acids with thesequence provided in or complementary to the GenBank entry referenced incolumn 3 of Table 2, which correspond to nucleic acid sequences setforth as SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ IDNO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27,SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO:37, SEQ ID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ IDNO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65,SEQ ID NO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73 SEQ ID NO:75, SEQ ID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ IDNO: 85, SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103,SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ IDNO: 113, SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, or SEQ ID NO:121, as well as functionally active fragments, derivatives, or orthologsthereof. An IMQ nucleic acid of this disclosure may be DNA, derived fromgenomic DNA or cDNA, or RNA. Genomic sequences of the genes listed inTable 2 are known and available in public databases such as GenBank.

In one embodiment, a functionally active IMQ nucleic acid encodes or iscomplementary to a nucleic acid that encodes a functionally active IMQpolypeptide. A functionally active IMQ nucleic acid also includesgenomic DNA that serves as a template for a primary RNA transcript(i.e., an mRNA precursor) that requires processing, such as splicing,before encoding the functionally active IMQ polypeptide. An IMQ nucleicacid can include other non-coding sequences, which may or may not betranscribed; such sequences include 5′ and 3′ UTRs, polyadenylationsignals and regulatory sequences that control gene expression, amongothers, as are known in the art. Some polypeptides require processingevents, such as proteolytic cleavage, covalent modification, etc., inorder to become fully active. Accordingly, functionally active nucleicacids may encode the mature or the pre-processed IMQ polypeptide, or anintermediate form. An IMQ polynucleotide can also include heterologouscoding sequences, for example, sequences that encode a marker includedto facilitate the purification of the fused polypeptide, or atransformation marker. In another embodiment, a functionally active IMQnucleic acid is capable of being used in the generation ofloss-of-function IMQ phenotypes, for instance, via antisensesuppression, co-suppression, etc. The disclosure also provides feed,meal, grain, food, or seed comprising a nucleic acid sequence thatencodes an IMQ polypeptide.

In one preferred embodiment, an IMQ nucleic acid used in the disclosedmethods comprises a nucleic acid sequence that encodes, or iscomplementary to a sequence that encodes, an IMQ polypeptide having atleast 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequenceidentity to a disclosed IMQ polypeptide sequence, for example the aminoacid sequence set forth as SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16,SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO:26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ IDNO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54,SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO:64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ IDNO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92,SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO:102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO:120, or SEQ ID NO: 122.

In another embodiment, an IMQ polypeptide comprises a polypeptidesequence with at least 50% or 60% identity to a disclosed IMQpolypeptide sequence (for example, the amino acid sequence set forth asSEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10,SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ IDNO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48,SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO:58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ IDNO: 68, SEQ ID NO: 70, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQID NO: 78, SEQ ID NO: 80, SEQ ID NO: 82, SEQ ID NO: 84, SEQ ID NO: 86,SEQ ID NO: 88, SEQ ID NO: 90, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO:96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQID NO: 106, SEQ ID NO: 108, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO:114, SEQ ID NO: 116, SEQ ID NO: 118, SEQ ID NO: 120, or SEQ ID NO: 122)and may have at least 70%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequenceidentity to a disclosed IMQ polypeptide sequence. In a furtherembodiment, an IMQ polypeptide comprises 50%, 60%, 70%, 80%, 85%, 90%,95%, 97%, 98%, or 99% sequence identity to a disclosed IMQ polypeptidesequence, and may include a conserved protein domain of the IMQpolypeptide (such as the protein domain(s) listed in column 8 of Table2). In another embodiment, an IMQ polypeptide comprises a polypeptidesequence with at least 50%, 60%, 70%, 80%, 85%, 90%, 95%, 97%, 98%, or99% sequence identity to a functionally active fragment of thepolypeptide referenced in column 5 of Table 2. In yet anotherembodiment, an IMQ polypeptide comprises a polypeptide sequence with atleast 50%, 60%, 70%, 80%, 90%, 95%, 97%, 98%, or 99% identity to thepolypeptide sequence of the GenBank entry referenced in column 5 ofTable 2 over its entire length and comprises a conserved proteindomain(s) listed in column 8 of Table 2.

In another aspect, an IMQ polynucleotide sequence is at least 50% to 60%identical over its entire length to a disclosed IMQ nucleic acidsequence, such as the nucleic acid sequence set forth as SEQ ID NO: 1,SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11,SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO:21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29, SEQ IDNO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQ ID NO: 39, SEQID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49,SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO: 57, SEQ ID NO:59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, SEQ IDNO: 69, SEQ ID NO: 71, SEQ ID NO: 73 SEQ ID NO: 75, SEQ ID NO: 77, SEQID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85, SEQ ID NO: 87,SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO: 95, SEQ ID NO:97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113, SEQ ID NO:115, SEQ ID NO: 117, SEQ ID NO: 119, or SEQ ID NO: 121, or nucleic acidsequences that are complementary to such an IMQ sequence, and maycomprise at least 70%, 80%, 85%, 90%, 95%, 97%, 98%, or 99% sequenceidentity to the disclosed IMQ sequence, or a functionally activefragment thereof, or complementary sequences. In another embodiment, adisclosed IMQ nucleic acid comprises a nucleic acid sequence as shown inSEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9,SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO:19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ IDNO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47,SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO:57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ IDNO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73 SEQ ID NO: 75, SEQID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85,SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO:95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ IDNO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113,SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, or SEQ ID NO: 121, ornucleic acid sequences that are complementary to such an IMQ sequence,and nucleic acid sequences that have substantial sequence homology to asuch IMQ sequences. As used herein, the phrase “substantial sequencehomology” refers to those nucleic acid sequences that have slight orinconsequential sequence variations from such IMQ sequences, i.e., thesequences function in substantially the same manner and encode an IMQpolypeptide.

As used herein, “percent (%) sequence identity” with respect to aspecified subject sequence, or a specified portion thereof, is definedas the percentage of nucleotides or amino acids in an identifiedsequence identical with the nucleotides or amino acids in the subjectsequence (or specified portion thereof), after aligning the sequencesand introducing gaps, if necessary to achieve the maximum percentsequence identity, as generated by the program WU-BLAST-2.0a19 (Altschulet al., J. Mol. Biol., 1990, 215:403-410) with search parameters set todefault values. The HSP S and HSP S2 parameters are dynamic values andare established by the program itself depending upon the composition ofthe particular sequence and composition of the particular databaseagainst which the sequence of interest is being searched. A “percent (%)identity value” is determined by the number of matching identicalnucleotides or amino acids divided by the sequence length for which thepercent identity is being reported. “Percent (%) amino acid sequencesimilarity” is determined by performing the same calculation as fordetermining % amino acid sequence identity, but including conservativeamino acid substitutions in addition to identical amino acids in thecomputation. A conservative amino acid substitution is one in which anamino acid is substituted for another amino acid having similarproperties such that the folding or activity of the protein is notsignificantly affected. Aromatic amino acids that can be substituted foreach other are phenylalanine, tryptophan, and tyrosine; interchangeablehydrophobic amino acids are leucine, isoleucine, methionine, and valine;interchangeable polar amino acids are glutamine and asparagine;interchangeable basic amino acids are arginine, lysine and histidine;interchangeable acidic amino acids are aspartic acid and glutamic acid;and interchangeable small amino acids are alanine, serine, threonine,cysteine and glycine.

Derivative nucleic acid molecules of the subject nucleic acid moleculesinclude sequences that selectively hybridize to the disclosed IMQnucleic acid sequences (for example, the nucleic acid sequence set forthas SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9,SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO:19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ IDNO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47,SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO:57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ IDNO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73 SEQ ID NO: 75, SEQID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85,SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO:95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ IDNO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113,SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, or SEQ ID NO: 121). Thestringency of hybridization can be controlled by temperature, ionicstrength, pH, and the presence of denaturing agents such as formamideduring hybridization and washing. Conditions routinely used are wellknown (see, e.g., Current Protocol in Molecular Biology, Vol. 1, Chap.2.10, John Wiley & Sons, Publishers (1994); Sambrook et al., 1989,Molecular Cloning: A Laboratory Manual (Second Edition), Cold SpringHarbor Press, Plainview, N.Y.).

In some embodiments, a nucleic acid molecule of the disclosure iscapable of hybridizing to a nucleic acid molecule containing thedisclosed nucleotide sequence under stringent hybridization conditionsthat are: prehybridization of filters containing nucleic acid for 8hours to overnight at 65° C. in a solution comprising 6× single strengthcitrate (SSC) (1×SSC is 0.15 M NaCl, 0.015 M Na citrate; pH 7.0),5×Denhardt's solution, 0.05% sodium pyrophosphate and 100 μg/ml herringsperm DNA; hybridization for 18-20 hours at 65° C. in a solutioncontaining 6×SSC, 1×Denhardt's solution, 100 μg/ml yeast tRNA and 0.05%sodium pyrophosphate; and washing of filters at 65° C. for 1 h in asolution containing 0.1×SSC and 0.1% SDS (sodium dodecyl sulfate). Inother embodiments, moderately stringent hybridization conditions areused that are: pretreatment of filters containing nucleic acid for 6 hat 40° C. in a solution containing 35% formamide, 5×SSC, 50 mM Tris-HCl(pH 7.5), 5 mM EDTA, 0.1% PVP, 0.1% Ficoll, 1% BSA, and 500 μg/mldenatured salmon sperm DNA; hybridization for 18-20 h at 40° C. in asolution containing 35% formamide, 5×SSC, 50 mM Tris-HCl (pH 7.5), 5 mMEDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 μg/ml salmon sperm DNA, and10% (wt/vol) dextran sulfate; followed by washing twice for 1 hour at55° C. in a solution containing 2×SSC and 0.1% SDS. Alternatively, lowstringency conditions can be used that comprise: incubation for 8 hoursto overnight at 37° C. in a solution comprising 20% formamide, 5×SSC, 50mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextransulfate, and 20 μg/ml denatured sheared salmon sperm DNA; hybridizationin the same buffer for 18 to 20 hours; and washing of filters in 1×SSCat about 37° C. for 1 hour.

As a result of the degeneracy of the genetic code, a number ofpolynucleotide sequences encoding an IMQ polypeptide can be produced.For example, codons may be selected to increase the rate at whichexpression of the polypeptide occurs in a particular host species, inaccordance with the optimum codon usage dictated by the particular hostorganism (see, e.g., Nakamura et al., 1999, Nucleic Acids Res. 27:292).Such sequence variants may be used in the methods disclosed herein.

The disclosed methods may use orthologs of a disclosed Arabidopsis IMQnucleic acid sequence. Representative putative orthologs of each of thedisclosed Arabidopsis IMQ genes are identified in column 3 of Table 3,below. Methods of identifying the orthologs in other plant species areknown in the art. In general, orthologs in different species retain thesame function, due to presence of one or more protein motifs and/or3-dimensional structures. In evolution, when a gene duplication eventfollows speciation, a single gene in one species, such as Arabidopsis,may correspond to multiple genes (paralogs) in another. As used herein,the term “orthologs” encompasses paralogs. When sequence data isavailable for a particular plant species, orthologs are generallyidentified by sequence homology analysis, such as BLAST analysis,usually using protein bait sequences. Sequences are assigned as apotential ortholog if the best hit sequence from the forward BLASTresult retrieves the original query sequence in the reverse BLAST(Huynen M A and Bork P, 1998, Proc. Natl. Acad. Sci., 95:5849-5856;Huynen M A et al., 2000, Genome Research, 10:1204-1210).

Programs for multiple sequence alignment, such as CLUSTAL (Thompson J Det al., 1994, Nucleic Acids Res. 22:4673-4680) may be used to highlightconserved regions and/or residues of orthologous proteins and togenerate phylogenetic trees. In a phylogenetic tree representingmultiple homologous sequences from diverse species (e.g., retrievedthrough BLAST analysis), orthologous sequences from two speciesgenerally appear closest on the tree with respect to all other sequencesfrom these two species. Structural threading or other analysis ofprotein folding (e.g., using software by ProCeryon, Biosciences,Salzburg, Austria) may also identify potential orthologs. Nucleic acidhybridization methods may also be used to find orthologous genes and arepreferred when sequence data are not available. Degenerate PCR andscreening of cDNA or genomic DNA libraries are common methods forfinding related gene sequences and are well known in the art (see, e.g.,Sambrook, 1989, Molecular Cloning: A Laboratory Manual (Second Edition),Cold Spring Harbor Press, Plainview, N.Y.; Dieffenbach and Dveksler,1995, PCR Primer: A Laboratory Manual, Cold Spring Harbor LaboratoryPress, NY). For instance, methods for generating a cDNA library from theplant species of interest and probing the library with partiallyhomologous gene probes are described in Sambrook et al. A highlyconserved portion of the Arabidopsis IMQ coding sequence may be used asa probe. IMQ ortholog nucleic acids may hybridize to the nucleic acid ofSEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9,SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO:19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ IDNO: 29, SEQ ID NO: 31, SEQ ID NO: 33, SEQ ID NO: 35, SEQ ID NO: 37, SEQID NO: 39, SEQ ID NO: 41, SEQ ID NO: 43, SEQ ID NO: 45, SEQ ID NO: 47,SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, SEQ ID NO: 55, SEQ ID NO:57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ IDNO: 67, SEQ ID NO: 69, SEQ ID NO: 71, SEQ ID NO: 73 SEQ ID NO: 75, SEQID NO: 77, SEQ ID NO: 79, SEQ ID NO: 81, SEQ ID NO: 83, SEQ ID NO: 85,SEQ ID NO: 87, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 93, SEQ ID NO:95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ IDNO: 105, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 111, SEQ ID NO: 113,SEQ ID NO: 115, SEQ ID NO: 117, SEQ ID NO: 119, or SEQ ID NO: 121 underhigh, moderate, or low stringency conditions. After amplification orisolation of a segment of a putative ortholog, that segment may becloned and sequenced by standard techniques and utilized as a probe toisolate a complete cDNA or genomic DNA clone.

Alternatively, it is possible to initiate an EST project to generate adatabase of sequence information for the plant species of interest. Inanother approach, antibodies that specifically bind known IMQpolypeptides are used for ortholog isolation (see, e.g., Harlow andLane, 1988, 1999, Antibodies: A Laboratory Manual, Cold Spring HarborLaboratory Press, NY). Western blot analysis can determine that an IMQortholog (i.e., a protein orthologous to a disclosed IMQ polypeptide) ispresent in a crude extract of a particular plant species. Whenreactivity is observed, the sequence encoding the candidate ortholog maybe isolated by screening expression libraries representing theparticular plant species. Expression libraries can be constructed in avariety of commercially available vectors, including lambda gt11, asdescribed in Sambrook, et al., 1989. Once the candidate ortholog(s) areidentified by any of these means, candidate orthologous sequence areused as bait (the “query”) for the reverse BLAST against sequences fromArabidopsis or other species in which IMQ nucleic acid and/orpolypeptide sequences have been identified.

IMQ nucleic acids and polypeptides may be obtained using any availablemethod. For instance, techniques for isolating cDNA or genomic DNAsequences of interest by screening DNA libraries or by using polymerasechain reaction (PCR), as previously described, are well known in theart. Alternatively, nucleic acid sequence may be synthesized. Any knownmethod, such as site directed mutagenesis (Kunkel T A et al., 1991,Methods Enzymol. 204:125-39), may be used to introduce desired changesinto a cloned nucleic acid.

In general, the methods disclosed herein involve incorporating thedesired form of the IMQ nucleic acid into a plant expression vector fortransformation of plant cells, and the IMQ polypeptide is expressed inthe host plant. Transformed plants and plant cells expressing an IMQpolypeptide express an IMQ phenotype and/or an IOQ phenotype and, in onespecific, non-limiting example, may have high (increased) oil, high(increased) protein, and/or low (decreased) fiber content.

An “isolated” IMQ nucleic acid molecule is other than in the form orsetting in which it is found in nature, and is identified and separatedfrom least one contaminant nucleic acid molecule with which it isordinarily associated in the natural source of the IMQ nucleic acid.However, an isolated IMQ nucleic acid molecule includes IMQ nucleic acidmolecules contained in cells that ordinarily express IMQ where, forexample, the nucleic acid molecule is in a chromosomal locationdifferent from that of natural cells.

Generation of Genetically Modified Plants with an Improved Oil QuantityPhenotype and/or an Improved Meal Quality Phenotype

The disclosed IMQ nucleic acids and polypeptides may be used in thegeneration of transgenic plants having a modified or altered oil,protein, and/or fiber content phenotype. As used herein, an “altered oilcontent (phenotype)” may refer to altered oil content in any part of theplant. In a preferred embodiment, altered expression of the IMQ gene ina plant is used to generate plants with a high oil content (phenotype).As used herein, an “altered protein content (phenotype)” may refer toaltered protein content in any part of the plant. In a preferredembodiment, altered expression of the IMQ gene in a plant is used togenerate plants with a high (or increased) protein content (phenotype).As used herein, an “altered fiber content (phenotype)” may refer toaltered fiber content in any part of the plant. In a preferredembodiment, altered expression of the IMQ gene in a plant is used togenerate plants with a low (or decreased) fiber content (phenotype). Thealtered oil, protein, and/or fiber content is often observed in seeds.Examples of a transgenic plant include plants comprising a planttransformation vector with a nucleotide sequence that encodes or iscomplementary to a sequence that encodes an IMQ polypeptide having theamino acid sequence as set forth in SEQ ID NO: 2, SEQ ID NO: 4, SEQ IDNO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ IDNO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34,SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO:44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ IDNO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 72,SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 78, SEQ ID NO: 80, SEQ ID NO:82, SEQ ID NO: 84, SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 90, SEQ IDNO: 92, SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, SEQ ID NO:110, SEQ ID NO: 112, SEQ ID NO: 114, SEQ ID NO: 116, SEQ ID NO: 118, SEQID NO: 120, or SEQ ID NO: 122, or an ortholog thereof.

Transgenic plants, such as corn, soybean and canola containing thedisclosed nucleic acid sequences, can be used in the production ofvegetable oil and meal. Vegetable oil is used in a variety of foodproducts, while meal from seed is used as an animal feed. Afterharvesting seed from transgenic plants, the seed is cleaned to removeplant stalks and other material and then flaked in roller mills to breakthe hulls. The crushed seed is heated to 75-100° C. to denaturehydrolytic enzymes, lyse the unbroken oil containing cells, and allowsmall oil droplets to coalesce. Most of the oil is then removed (and canbe recovered) by pressing the seed material in a screw press. Theremaining oil is removed from the presscake by extraction with andorganic solvents, such as hexane. The solvent is removed from the mealby heating it to approximately 100° C. After drying, the meal is thengranulated to a consistent form. The meal, containing the protein,digestible carbohydrate, and fiber of the seed, may be mixed with othermaterials prior to being used as an animal feed.

The methods described herein for generating transgenic plants aregenerally applicable to all plants. Although activation tagging and geneidentification is carried out in Arabidopsis, the IMQ nucleic acidsequence (or an ortholog, variant or fragment thereof) may be expressedin any type of plant. In a preferred embodiment, oil-producing plantsproduce and store triacylglycerol in specific organs, primarily inseeds. Such species include soybean (Glycine max), rapeseed and canola(including Brassica napus, B. campestris), sunflower (Helianthus annus),cotton (Gossypium hirsutum), corn (Zea mays), cocoa (Theobroma cacao),safflower (Carthamus tinctorius), oil palm (Elaeis guineensis), coconutpalm (Cocos nucifera), flax (Linum usitatissimum), castor (Ricinuscommunis), and peanut (Arachis hypogaea), as well as wheat, rice andoat. Fruit- and vegetable-bearing plants, grain-producing plants,nut-producing plants, rapid cycling Brassica species, alfalfa (Medicagosativa), tobacco (Nicotiana), turfgrass (Poaceae family), other foragecrops, and wild species may also be a source of unique fatty acids. Inother embodiments, any plant expressing the IMQ nucleic acid sequencecan also express increased protein and/or decreased fiber content in aspecific plant part or organ, such as in seeds.

The skilled artisan will recognize that a wide variety of transformationtechniques exist in the art, and new techniques are continually becomingavailable. Any technique that is suitable for the target host plant canbe employed within the scope of the present invention. For example, theconstructs can be introduced in a variety of forms including, but notlimited to, as a strand of DNA, in a plasmid, or in an artificialchromosome. The introduction of the constructs into the target plantcells can be accomplished by a variety of techniques, including, but notlimited to, Agrobacterium-mediated transformation, electroporation,microinjection, microprojectile bombardment, calcium-phosphate-DNAco-precipitation, or liposome-mediated transformation of a heterologousnucleic acid. The transformation of the plant is preferably permanent,i.e. by integration of the introduced expression constructs into thehost plant genome, so that the introduced constructs are passed ontosuccessive plant generations. Depending upon the intended use, aheterologous nucleic acid construct comprising an IMQ polynucleotide mayencode the entire protein or a biologically active portion thereof.

In one embodiment, binary Ti-based vector systems may be used totransfer polynucleotides. Standard Agrobacterium binary vectors areknown to those of skill in the art, and many are commercially available(e.g., pBI121 Clontech Laboratories, Palo Alto, Calif.). A construct orvector may include a plant promoter to express the nucleic acid moleculeof choice. In a preferred embodiment, the promoter is a plant promoter.

The optimal procedure for transformation of plants with Agrobacteriumvectors will vary with the type of plant being transformed. Exemplarymethods for Agrobacterium-mediated transformation include transformationof explants of hypocotyl, shoot tip, stem or leaf tissue, derived fromsterile seedlings and/or plantlets. Such transformed plants may bereproduced sexually, or by cell or tissue culture. Agrobacteriumtransformation has been previously described for a large number ofdifferent types of plants and methods for such transformation may befound in the scientific literature. Of particular relevance are methodsto transform commercially important crops, such as plants of theBrassica species, including canola and rapeseed, (De Block et al., 1989,Plant Physiol., 91:694-701), sunflower (Everett et al., 1987,Bio/Technology, 5:1201), soybean (Christou et al., 1989, Proc. Natl.Acad. Sci USA, 86:7500-7504; Kline et al., 1987, Nature, 327:70), wheat,rice and oat.

Expression (including transcription and translation) of an IMQ nucleicacid sequence may be regulated with respect to the level of expression,the tissue type(s) where expression takes place and/or developmentalstage of expression. A number of heterologous regulatory sequences(e.g., promoters and enhancers) are available for controlling theexpression of an IMQ nucleic acid. These include constitutive, inducibleand regulatable promoters, as well as promoters and enhancers thatcontrol expression in a tissue- or temporal-specific manner. Exemplaryconstitutive promoters include the raspberry E4 promoter (U.S. Pat. Nos.5,783,393 and 5,783,394), the nopaline synthase (NOS) promoter (Ebert etal., Proc. Natl. Acad. Sci. (U.S.A.) 84:5745-5749, 1987), the octopinesynthase (OCS) promoter (which is carried on tumor-inducing plasmids ofAgrobacterium tumefaciens), the caulimovirus promoters such as thecauliflower mosaic virus (CaMV) 19S promoter (Lawton et al., Plant Mol.Biol. 9:315-324, 1987) and the CaMV 35S promoter (Odell et al., Nature313:810-812, 1985 and Jones J D et al, 1992, Transgenic Res.,1:285-297), the figwort mosaic virus 35S-promoter (U.S. Pat. No.5,378,619), the light-inducible promoter from the small subunit ofribulose-1,5-bis-phosphate carboxylase (ssRUBISCO), the Adh promoter(Walker et al., Proc. Natl. Acad. Sci. (U.S.A.) 84:6624-6628, 1987), thesucrose synthase promoter (Yang et al., Proc. Natl. Acad. Sci. (U.S.A.)87:4144-4148, 1990), the R gene complex promoter (Chandler et al., ThePlant Cell 1:1175-1183, 1989), the chlorophyll a/b binding protein genepromoter, the CsVMV promoter (Verdaguer et al., 1998, Plant Mol Biol.,37:1055-1067), and the melon actin promoter (published PCT applicationWO0056863). Exemplary tissue-specific promoters include the tomato E4and E8 promoters (U.S. Pat. No. 5,859,330) and the tomato 2AII genepromoter (Van Haaren et al., 1993, Plant Mol Bio., 21:625-640).

In one preferred embodiment, expression of the IMQ nucleic acid sequenceis under control of regulatory sequences from genes whose expression isassociated with early seed and/or embryo development. Indeed, in apreferred embodiment, the promoter used is a seed-enhanced promoter.Examples of such promoters include the 5′ regulatory regions from suchgenes as napin (Kridl et al., Seed Sci. Res. 1:209:219, 1991), globulin(Belanger and Kriz, Genet., 129: 863-872, 1991, GenBank Accession No.L22295), gamma zein Z 27 (Lopes et al., Mol Gen Genet., 247:603-613,1995), L3 oleosin promoter (U.S. Pat. No. 6,433,252), phaseolin (Bustoset al., Plant Cell, 1(9):839-853, 1989), arcelin5 (U.S. Application No.2003/0046727), a soybean 7S promoter, a 7Sα promoter (U.S. ApplicationNo. 2003/0093828), the soybean 7Sα′ beta conglycinin promoter, a 7S a′promoter (Beachy et al., EMBO J., 4:3047, 1985; Schuler et al., NucleicAcid Res., 10(24):8225-8244, 1982), soybean trypsin inhibitor (Riggs etal., Plant Cell 1(6):609-621, 1989), ACP (Baerson et al., Plant Mol.Biol., 22(2):255-267, 1993), stearoyl-ACP desaturase (Slocombe et al.,Plant Physiol. 104(4):167-176, 1994), soybean a′ subunit ofβ-conglycinin (Chen et al., Proc. Natl. Acad. Sci. 83:8560-8564, 1986),Vicia faba USP (P-Vf.Usp, SEQ ID NO: 1, 2, and 3 in (U.S. ApplicationNo. 2003/229918) and Zea mays L3 oleosin promoter (Hong et al., PlantMol. Biol., 34(3):549-555, 1997). Also included are the zeins, which area group of storage proteins found in corn endosperm. Genomic clones forzein genes have been isolated (Pedersen et al., Cell, 29:1015-1026,1982; and Russell et al., Transgenic Res. 6(2):157-168) and thepromoters from these clones, including the 15 kD, 16 kD, 19 kD, 22 kD,27 kD and genes, could also be used. Other promoters known to function,for example, in corn include the promoters for the following genes:waxy, Brittle, Shrunken 2, Branching enzymes I and II, starch synthases,debranching enzymes, oleosins, glutelins and sucrose synthases. Legumegenes whose promoters are associated with early seed and embryodevelopment include V. faba legumin (Baumlein et al., 1991, Mol. Gen.Genet. 225:121-8; Baumlein et al., 1992, Plant J. 2:233-9), V. faba usp(Fiedler et al., 1993, Plant Mol. Biol. 22:669-79), pea convicilin (Bownet al., 1988, Biochem. J. 251:717-26), pea lectin (dePater et al., 1993,Plant Cell 5:877-86), P. vulgaris beta phaseolin (Bustos et al., 1991,EMBO J. 10:1469-79), P. vulgaris DLEC2 and PHS [beta] (Bobb et al.,1997, Nucleic Acids Res. 25:641-7), and soybean beta-Conglycinin, 7Sstorage protein (Chamberland et al., 1992, Plant Mol. Biol. 19:937-49).

Cereal genes whose promoters are associated with early seed and embryodevelopment include rice glutelin (“GluA-3,” Yoshihara and Takaiwa,1996, Plant Cell Physiol. 37:107-11; “GluB-1,” Takaiwa et al., 1996,Plant Mol. Biol. 30:1207-21; Washida et al., 1999, Plant Mol. Biol.40:1-12; “Gt3,” Leisy et al., 1990, Plant Mol. Biol. 14:41-50), riceprolamin (Zhou & Fan, 1993, Transgenic Res. 2:141-6), wheat prolamin(Hammond-Kosack et al., 1993, EMBO J. 12:545-54), maize zein (Z4, Matzkeet al., 1990, Plant Mol. Biol. 14:323-32), and barley B-hordeins(Entwistle et al., 1991, Plant Mol. Biol. 17:1217-31).

Other genes whose promoters are associated with early seed and embryodevelopment include oil palm GLO7A (7S globulin, Morcillo et al., 2001,Physiol. Plant 112:233-243), Brassica napus napin, 2S storage protein,and napA gene (Josefsson et al., 1987, J. Biol. Chem. 262:12196-201;Stalberg et al., 1993, Plant Mol. Biol. 1993 23:671-83; Ellerstrom etal., 1996, Plant Mol. Biol. 32:1019-27), Brassica napus oleosin (Keddieet al., 1994, Plant Mol. Biol. 24:327-40), Arabidopsis oleosin (Plant etal., 1994, Plant Mol. Biol. 25:193-205), Arabidopsis FAE1 (Rossak etal., 2001, Plant Mol. Biol. 46:717-25), Canavalia gladiata conA(Yamamoto et al., 1995, Plant Mol. Biol. 27:729-41), and Catharanthusroseus strictosidine synthase (Str, Ouwerkerk and Memelink, 1999, Mol.Gen. Genet. 261:635-43). In another preferred embodiment, regulatorysequences from genes expressed during oil biosynthesis are used (see,e.g., U.S. Pat. No. 5,952,544). Alternative promoters are from plantstorage protein genes (Bevan et al., 1993, Philos. Trans. R. Soc. Lond.B. Biol. Sci. 342:209-15). Additional promoters that may be utilized aredescribed, for example, in U.S. Pat. Nos. 5,378,619; 5,391,725;5,428,147; 5,447,858; 5,608,144; 5,608,144; 5,614,399; 5,633,441;5,633,435; and 4,633,436.

In yet another aspect, in some cases it may be desirable to inhibit theexpression of the endogenous IMQ nucleic acid sequence in a host cell.Exemplary methods for practicing this aspect of the invention include,but are not limited to antisense suppression (Smith, et al., 1988,Nature, 334:724-726; van der Krol et al., 1988, BioTechniques,6:958-976); co-suppression (Napoli, et al., 1990, Plant Cell,2:279-289); ribozymes (PCT Publication WO 97/10328); and combinations ofsense and antisense (Waterhouse, et al., 1998, Proc. Natl. Acad. Sci.USA, 95:13959-13964). Methods for the suppression of endogenoussequences in a host cell typically employ the transcription ortranscription and translation of at least a portion of the sequence tobe suppressed. Such sequences may be homologous to coding as well asnon-coding regions of the endogenous sequence. Antisense inhibition mayuse the entire cDNA sequence (Sheehy et al., 1988, Proc. Natl. Acad.Sci. USA, 85:8805-8809), a partial cDNA sequence including fragments of5′ coding sequence, (Cannon et al., 1990, Plant Mol. Biol., 15:39-47),or 3′ non-coding sequences (Ch′ng et al., 1989, Proc. Natl. Acad. Sci.USA, 86:10006-10010). Cosuppression techniques may use the entire cDNAsequence (Napoli et al., 1990, Plant Cell, 2:279-289; van der Krol etal., 1990, Plant Cell, 2:291-299), or a partial cDNA sequence (Smith etal., 1990, Mol. Gen. Genetics, 224:477-481).

Standard molecular and genetic tests may be performed to further analyzethe association between a nucleic acid sequence and an observedphenotype. Exemplary techniques are described below.

1. DNA/RNA Analysis

The stage- and tissue-specific gene expression patterns in mutant versuswild-type lines may be determined, for instance, by in situhybridization. Analysis of the methylation status of the gene,especially flanking regulatory regions, may be performed. Other suitabletechniques include over-expression, ectopic expression, expression inother plant species and gene knock-out (reverse genetics, targetedknock-out, viral induced gene silencing (VIGS; see, Baulcombe D, 1999,Arch. Virol. Suppl. 15:189-201).

In a preferred application expression profiling, generally by microarrayanalysis, is used to simultaneously measure differences or inducedchanges in the expression of many different genes. Techniques formicroarray analysis are well known in the art (Schena M et al., Science1995 270:467-470; Baldwin D et al., 1999, Cur. Opin. Plant Biol.2(2):96-103; Dangond F, Physiol Genomics (2000) 2:53-58; van Hal N L etal., J Biotechnol. (2000) 78:271-280; Richmond T and Somerville S, Curr.Opin. Plant Biol. 2000 3:108-116). Expression profiling of individualtagged lines may be performed. Such analysis can identify other genesthat are coordinately regulated as a consequence of the over-expressionof the gene of interest, which may help to place an unknown gene in aparticular pathway.

2. Gene Product Analysis

Analysis of gene products may include recombinant protein expression,antisera production, immunolocalization, biochemical assays forcatalytic or other activity, analysis of phosphorylation status, andanalysis of interaction with other proteins via yeast two-hybrid assays.

3. Pathway Analysis

Pathway analysis may include placing a gene or gene product within aparticular biochemical, metabolic or signaling pathway based on itsmis-expression phenotype or by sequence homology with related genes.Alternatively, analysis may comprise genetic crosses with wild-typelines and other mutant lines (creating double mutants) to order the genein a pathway, or determining the effect of a mutation on expression ofdownstream “reporter” genes in a pathway.

Generation of Mutated Plants with an Improved Oil Quantity Phenotypeand/or Improved Meal Quality Phenotype

Additional methods are disclosed herein of generating a plant having anIMQ and/or an IOQ phenotype, wherein a plant is identified that has anallele in its IMQ nucleic acid sequence that results in an IMQ phenotypeand/or an IOQ phenotype, compared to plants lacking the allele. Theplant can generate progeny, wherein the progeny inherit the allele andhave an IMQ phenotype and/or an IOQ phenotype. For example, providedherein is a method of identifying plants that have mutations in theendogenous IMQ nucleic acid sequence that confer an IMQ phenotype and/oran IOQ phenotype and generating progeny of these plants with an IMQand/or IOQ phenotype that are not genetically modified. In someembodiments, the plants have an IMQ phenotype with an altered proteinand/or fiber content or seed meal content, or an IOQ phenotype, with analtered oil content.

In one method, called “TILLING” (for targeting induced local lesions ingenomes), mutations are induced in the seed of a plant of interest, forexample, using EMS (ethylmethane sulfonate) treatment. The resultingplants are grown and self-fertilized, and the progeny are used toprepare DNA samples. PCR amplification and sequencing of the IMQ nucleicacid sequence is used to identify whether a mutated plant has a mutationin the IMQ nucleic acid sequence. Plants having IMQ mutations may thenbe tested for altered oil, protein, and/or fiber content, oralternatively, plants may be tested for altered oil, protein, and/orfiber content, and then PCR amplification and sequencing of the IMQnucleic acid sequence is used to determine whether a plant havingaltered oil, protein, and/or fiber content has a mutated IMQ nucleicacid sequence. TILLING can identify mutations that may alter theexpression of specific genes or the activity of proteins encoded bythese genes (see Colbert et al., 2001, Plant Physiol. 126:480-484;McCallum et al., 2000, Nature Biotechnology 18:455-457).

In another method, a candidate gene/Quantitative Trait Locus (QTLs)approach can be used in a marker-assisted breeding program to identifyalleles of or mutations in the IMQ nucleic acid sequence or orthologs ofthe IMQ nucleic acid sequence that may confer altered oil, protein,and/or fiber content (see Bert et al., Theor Appl Genet., 2003 June;107(1):181-9; and Lionneton et al., Genome, 2002 December;45(6):1203-15). Thus, in a further aspect of the disclosure, an IMQnucleic acid is used to identify whether a plant having altered oil,protein, and/or fiber content has a mutation an endogenous IMQ nucleicacid sequence or has a particular allele that causes altered oil,protein, and/or fiber content.

While the disclosure has been described with reference to specificmethods and embodiments, it will be appreciated that variousmodifications and changes may be made without departing from thedisclosure. All publications cited herein are expressly incorporatedherein by reference for the purpose of describing and disclosingcompositions and methodologies that might be used in connection with thedisclosure. All cited patents, patent applications, and sequenceinformation in referenced public databases are also incorporated byreference.

EXAMPLES Example 1 Generation of Plants with an IMQ Phenotype and/or anIOQ Phenotype by Transformation with an Activation Tagging Construct

This Example describes the generation of transgenic plants with alteredoil, protein, and/or fiber content.

Mutants were generated using the activation tagging “ACTTAG” vector,pSKI015 (GI#6537289; Weigel D et al., 2000, Plant Physiology,122:1003-1013). Standard methods were used for the generation ofArabidopsis transgenic plants, and were essentially as described inpublished application PCT WO0183697. Briefly, T0 Arabidopsis (Col-0)plants were transformed with Agrobacterium carrying the pSKI015 vector,which comprises T-DNA derived from the Agrobacterium Ti plasmid, anherbicide resistance selectable marker gene, and the 4×CaMV 35S enhancerelement. Transgenic plants were selected at the T1 generation based onherbicide resistance. T2 seed (from T1 plants) was harvested and sown insoil. T2 plants were exposed to the herbicide to kill plants lacking theACTTAG vector. T2 plants were grown to maturity, allowed toself-fertilize and set seed. T3 seed (from the T2 plants) was harvestedin bulk for each line.

T3 seed was analyzed by Near Infrared Spectroscopy (NIR) at the time ofharvest. NIR spectra were captured using a Bruker 22 near infraredspectrometer. Bruker Software was used to estimate total seed oil, totalseed protein and total seed fiber content using data from NIR analysisand reference methods according to the manufacturer's instructions. Oilcontent predicting calibrations were developed following the generalmethod of AOCS Procedure Am1-92, Official Methods and RecommendedPractices of the American Oil Chemists Society, 5th Ed., AOCS,Champaign, Ill. A NIR protein content predicting calibration wasdeveloped using total nitrogen content data of seed samples followingthe general method of Dumas Procedure AOAC 968.06 (Official Methods ofAnalysis of AOAC International 17^(th) Edition AOAC, Gaithersburg, Md.).A fiber content predicting calibration was developed by measuring crudefiber content in a set of seed samples. Fiber content of in a known massof seed was determined using the method of Honig and Rackis, (1979, J.Agri. Food Chem., 27: 1262-1266). Digestible protein content of in aknown mass of seed was determined by quantifying the individual aminoacids liberated by an acid hydrolysis Steine and Moore (1958, Anal.Chem., 30:1185-1190). The quantification was performed by the AminoQuant (Agilent). The undigested protein remaining associated with thenon-digestible fraction is measured by the same method described for thewhole seed homogenate. Digestible protein content is determined bysubtracting the amount of undigested protein associated with thenon-digestible fraction from the total amount of protein in the seedsample.

Seed oil, protein, digestible protein and fiber values in 82,274 lineswere determined by NIR spectroscopy and normalized to allow comparisonof seed component values in plants grown at different times. Oil,protein and fiber values were normalized by calculating the average oil,protein and fiber values in seed from all plants planted on the same day(including a large number of other ACTTAG plants, including control,wild-type, or non-transgenic plants). The seed components for each linewas expressed as a “percent relative value” which was calculated bydividing the component value for each line with the average componentvalue for all lines planted on the same day (which should approximatethe value in control, wild-type, or non-transgenic plants). The “percentrelative protein” and “percent relative fiber” were calculatedsimilarly.

Inverse PCR was used to recover genomic DNA flanking the T-DNAinsertion. The PCR product was subjected to sequence analysis and placedon the genome using a basic BLASTN search and/or a search of theArabidopsis Information Resource (TAIR) database (available at thepublicly available website). Promoters within 9 kb of the enhancers inthe ACTTAG element are considered to be within “activation space.” Geneswith T-DNA inserts within coding sequences were not considered to bewithin “activation space.” The ACTTAG lines with the above average oiland protein values, and below average fiber values were identified andare listed in column 3 of Table 1.

TABLE 1 4. 5. Relative Relative 6. Seed Seed Relative 1. Gene 3. ACTTAGProtein Fiber Seed Oil 7. alias 2. Tair Line Content Content Content GCFA IMQ66.4 At4g30810 W000092135 105.82% 89.32% 100.61% IMQ67.1 At4g32230W000148824 126.48% 88.73% 86.88% IMQ67.2 At4g32240 W000148824 126.48%88.73% 86.88% 93.73% IMQ67.3 At4g32250 W000148824 126.48% 88.73% 86.88%IMQ67.3 At4g32250 W000148824 126.48% 88.73% 86.88% IMQ67.3 At4g32250W000148824 126.48% 88.73% 86.88% IMQ67.4 At4g32260 W000148824 126.48%88.73% 86.88% IMQ68.1 At4g36560 W000203116 74.59% 97.16% 124.22% IMQ68.2At4g36570 W000203116 74.59% 97.16% 124.22% IMQ68.3 At4g36580 W00020311674.59% 97.16% 124.22% IMQ68.4 At4g36590 W000203116 74.59% 97.16% 124.22%IMQ68.5 At4g36600 W000203116 74.59% 97.16% 124.22% IMQ69.1 At4g38510W000050668 106.15% 91.34% 95.23% IMQ69.1 At4g38510 W000050668 106.15%91.34% 95.23% IMQ69.1 At4g38510 W000050668 106.15% 91.34% 95.23% IMQ69.1At4g38510 W000050668 106.15% 91.34% 95.23% IMQ69.2 At4g38520 W000050668106.15% 91.34% 95.23% IMQ69.2 At4g38520 W000050668 106.15% 91.34% 95.23%IMQ69.2 At4g38520 W000050668 106.15% 91.34% 95.23% IMQ69.3 At4g38530W000050668 106.15% 91.34% 95.23% 104.43% IMQ69.4 At4g38540 W000050668106.15% 91.34% 95.23% 104.43% IMQ69.5 At4g38550 W000050668 106.15%91.34% 95.23% IMQ69.6 At4g38560 W000050668 106.15% 91.34% 95.23% IMQ69.7At4g38570 W000050668 106.15% 91.34% 95.23% IMQ70.1 At5g11820 W000088626111.40% 91.87% 89.89% IMQ70.2 At5g11830 W000088626 111.40% 91.87% 89.89%IMQ70.3 At5g11840 W000088626 111.40% 91.87% 89.89% IMQ70.4 At5g11850W000088626 111.40% 91.87% 89.89% IMQ70.5 At5g11860 W000088626 111.40%91.87% 89.89% IMQ70.5 At5g11860 W000088626 111.40% 91.87% 89.89% IMQ70.5At5g11860 W000088626 111.40% 91.87% 89.89% IMQ70.6 At5g11870 W000088626111.40% 91.87% 89.89% IMQ70.7 At5g11880 W000088626 111.40% 91.87% 89.89%IMQ71.1 At5g12990 W000085787 140.67% 97.38% 76.10% IMQ71.2 At5g13000W000085787 140.67% 97.38% 76.10% 89.87% IMQ71.3 At5g13010 W000085787140.67% 97.38% 76.10% 89.87% IMQ72.1 At5g15700 W000114932 113.33% 94.87%96.37% IMQ72.2 At5g15710 W000114932 113.33% 94.87% 96.37% IMQ72.3At5g15720 W000114932 113.33% 94.87% 96.37% IMQ72.4 At5g15725 W000114932113.33% 94.87% 96.37% IMQ72.5 At5g15730 W000114932 113.33% 94.87% 96.37%IMQ73.1 At5g35960 W000154027 105.39% 92.83% 100.07% IMQ73.1 At5g35960W000192343 102.55% 87.48% 105.14% IMQ73.2 At5g35970 W000154027 105.39%92.83% 100.07% IMQ73.2 At5g35970 W000192343 102.55% 87.48% 105.14%IMQ73.3 At5g35980 W000154027 105.39% 92.83% 100.07% 103.52% IMQ73.3At5g35980 W000192343 102.55% 87.48% 105.14% IMQ73.3 At5g35980 W000154027105.39% 92.83% 100.07% IMQ73.3 At5g35980 W000192343 102.55% 87.48%105.14% IMQ74.1 At5g38530 W000095258 119.82% 93.87% 88.43% IMQ74.2At5g38540 W000095258 119.82% 93.87% 88.43% 98.68% IMQ74.3 At5g38550W000095258 119.82% 93.87% 88.43% IMQ74.4 At5g38560 W000095258 119.82%93.87% 88.43% IMQ75.1 At5g40300 W000209623 88.58% 86.86% 117.55% 121.36%IMQ75.2 At5g40310 W000209623 88.58% 86.86% 117.55% IMQ75.3 At5g40320W000209623 88.58% 86.86% 117.55% 121.36% IMQ76.1 At5g44180 W000064064118.55% 96.22% 85.55% IMQ77.1 At5g62600 W000156072 119.26% 92.93% 90.48%IMQ77.2 At5g62610 W000156072 119.26% 92.93% 90.48% IMQ78.1 At5g67580W000156087 111.66% 92.09% 96.86% IMQ78.1 At5g67580 W000156087 111.66%92.09% 96.86% IMQ78.2 At5g67590 W000156087 111.66% 92.09% 96.86% IMQ78.3At5g67600 W000156087 111.66% 92.09% 96.86% IMQ78.4 At5g67610 W000156087111.66% 92.09% 96.86% 101.88% IMQ78.4 At5g67610 W000156087 111.66%92.09% 96.86%

TABLE 2 7. Putative 5. biochemical 1. Gene 3. Nucleic 4. SEQ IDPolypeptide 6. SEQ ID function/protein 8. Conserved protein alias 2.Tair Acid seq. GI# NO seq. GI# NO name domain IMQ66.4 At4g30810gi|30688809 SEQ ID gi|18417667 SEQ ID SCPL29; catalytic/ IPR000379 NO: 1NO: 2 serine Esterase/lipase/thioesterase; carboxypeptidase IPR001563Peptidase S10, serine carboxypeptidase IMQ67.1 At4g32230 gi|18417967 SEQID gi|15236713 SEQ ID unknown protein IPR000719 Protein kinase NO: 3 NO:4 IMQ67.2 At4g32240 gi|42567326 SEQ ID gi|18417969 SEQ ID unknownprotein NO: 5 NO: 6 IMQ67.3 At4g32250 gi|79326107 SEQ ID gi|79326108 SEQID ATP binding/kinase/ IPR000719 Protein kinase; NO: 7 NO: 8 proteinkinase/ IPR008271 Serine/threonine protein protein kinase, active siteserine/threonine kinase/protein- tyrosine kinase IMQ67.3 At4g32250gi|30689315 SEQ ID gi|30689316 SEQ ID ATP binding/kinase/ IPR000719Protein kinase; NO: 9 NO: 10 protein kinase/ IPR008271 Serine/threonineprotein protein kinase, active site serine/threonine kinase/protein-tyrosine kinase IMQ67.3 At4g32250 gi|30689320 SEQ ID gi|22329080 SEQ IDATP binding/kinase/ IPR000719 Protein kinase; NO: 11 NO: 12 proteinkinase/ IPR008271 Serine/threonine protein protein kinase, active siteserine/threonine kinase/protein- tyrosine kinase IMQ67.4 At4g32260gi|30689323 SEQ ID gi|15236722 SEQ ID hydrolase, acting on IPR002146H+-transporting NO: 13 NO: 14 acid anhydrides, two-sector ATPase, B/B′catalyzing subunit; transmembrane IPR005864 ATP synthase F0, movement ofsubunit B substances IMQ68.1 At4g36560 gi|18419861 SEQ ID gi|15234443SEQ ID unknown protein NO: 15 NO: 16 IMQ68.2 At4g36570 gi|18419863 SEQID gi|15234454 SEQ ID DNA binding/ IPR001005 Myb, DNA-binding NO: 17 NO:18 transcription factor IMQ68.3 At4g36580 gi|18419864 SEQ ID gi|15234455SEQ ID ATP binding/ IPR000641 CbxX/CfqX; NO: 19 NO: 20ATPase/nucleoside- IPR003593 AAA ATPase; triphosphatase/ IPR003959 AAAATPase, nucleotide binding central region; IPR003960 AAA-proteinsubdomain IMQ68.4 At4g36590 gi|18419866 SEQ ID gi|15234456 SEQ ID DNAbinding/ IPR002100 Transcription NO: 21 NO: 22 transcription factorfactor, MADS-box IMQ68.5 At4g36600 gi|30690703 SEQ ID gi|30690704 SEQ IDunknown protein IPR004238 Late NO: 23 NO: 24 embryogenesis abundantprotein IMQ69.1 At4g38510 gi|79326467 SEQ ID gi|79326468 SEQ ID ATPbinding/ IPR000194 H+-transporting NO: 25 NO: 26 hydrogen-exportingtwo-sector ATPase, alpha/beta ATPase, subunit, central region;phosphorylative IPR000793 H+-transporting mechanism/ two-sector ATPase,alpha/beta hydrogen- subunit, C-terminal; transporting ATP IPR004100H+-transporting synthase, rotational two-sector ATPase, alpha/betamechanism/ subunit, N-terminal; hydrogen- IPR005723 ATP synthase V-transporting ATPase, type, B subunit rotational mechanism IMQ69.1At4g38510 gi|79326459 SEQ ID gi|79326460 SEQ ID ATP binding/ IPR000194H+-transporting NO: 27 NO: 28 hydrogen-exporting two-sector ATPase,alpha/beta ATPase, subunit, central region; phosphorylative IPR000793H+-transporting mechanism/ two-sector ATPase, alpha/beta hydrogen-subunit, C-terminal; transporting ATP IPR004100 H+-transportingsynthase, rotational two-sector ATPase, alpha/beta mechanism/ subunit,N-terminal; hydrogen- IPR005723 ATP synthase V- transporting ATPase,type, B subunit rotational mechanism IMQ69.1 At4g38510 gi|42573220 SEQID gi|42573221 SEQ ID ATP binding/ IPR000194 H+-transporting NO: 29 NO:30 hydrogen-exporting two-sector ATPase, alpha/beta ATPase, subunit,central region; phosphorylative IPR000793 H+-transporting mechanism/two-sector ATPase, alpha/beta hydrogen- subunit, C-terminal;transporting ATP IPR004100 H+-transporting synthase, rotationaltwo-sector ATPase, alpha/beta mechanism/ subunit, N-terminal; hydrogen-IPR005723 ATP synthase V- transporting ATPase, type, B subunitrotational mechanism IMQ69.1 At4g38510 gi|30691994 SEQ ID gi|15233891SEQ ID ATP binding/ IPR000194 H+-transporting NO: 31 NO: 32hydrogen-exporting two-sector ATPase, alpha/beta ATPase, subunit,central region; phosphorylative IPR000793 H+-transporting mechanism/two-sector ATPase, alpha/beta hydrogen- subunit, C-terminal;transporting ATP IPR004100 H+-transporting synthase, rotationaltwo-sector ATPase, alpha/beta mechanism/ subunit, N-terminal; hydrogen-IPR005723 ATP synthase V- transporting ATPase, type, B subunitrotational mechanism IMQ69.2 At4g38520 gi|42573222 SEQ ID gi|42573223SEQ ID protein phosphatase IPR000222 Protein NO: 33 NO: 34 2C familyprotein/ phosphatase 2C; PP2C family protein IPR001932 Proteinphosphatase 2C-like IMQ69.2 At4g38520 gi|42567510 SEQ ID gi|22329238 SEQID catalytic/protein IPR000222 Protein NO: 35 NO: 36 phosphatase type 2Cphosphatase 2C; IPR001932 Protein phosphatase 2C-like IMQ69.2 At4g38520gi|79313268 SEQ ID gi|79313269 SEQ ID catalytic/protein IPR000222Protein NO: 37 NO: 38 phosphatase type 2C phosphatase 2C; IPR001932Protein phosphatase 2C-like IMQ69.3 At4g38530 gi|79499921 SEQ IDgi|79499922 SEQ ID ATPLC1; IPR000008 C2; NO: 39 NO: 40 phospholipase CIPR000909 Phosphatidylinositol-specific phospholipase C, X region;IPR001192 Phosphoinositide- specific phospholipase C (PLC); IPR008973 C2calcium/lipid- binding region, CaLB; IPR001711Phosphatidylinositol-specific phospholipase C, Y domain; IMQ69.4At4g38540 gi|30692001 SEQ ID gi|15233923 SEQ ID monooxygenase IPR001327FAD-dependent NO: 41 NO: 42 pyridine nucleotide-disulphideoxidoreductase; IPR003042 Aromatic-ring hydroxylase; IPR006076 FADdependent oxidoreductase IMQ69.5 At4g38550 gi|42567511 SEQ IDgi|22329240 SEQ ID unknown protein IPR007942 Arabidopsis NO: 43 NO: 44phospholipase-like; IPR006706 Extensin-like region IMQ69.6 At4g38560gi|18420262 SEQ ID gi|15233931 SEQ ID unknown protein IPR007942Arabidopsis NO: 45 NO: 46 phospholipase-like IMQ69.7 At4g38570gi|30692008 SEQ ID gi|15233934 SEQ ID unknown protein IPR000462CDP-alcohol NO: 47 NO: 48 phosphatidyltransferase IMQ70.1 At5g11820gi|18416683 SEQ ID gi|15239789 SEQ ID unknown protein IPR010264 Plantself- NO: 49 NO: 50 incompatibility S1 IMQ70.2 At5g11830 gi|18416686 SEQID gi|15239794 SEQ ID unknown protein IPR010264 Plant self- NO: 51 NO:52 incompatibility S1 IMQ70.3 At5g11840 gi|18416688 SEQ ID gi|15239796SEQ ID unknown protein IPR009631 Protein of NO: 53 NO: 54 unknownfunction DUF1230 IMQ70.4 At5g11850 gi|30683823 SEQ ID gi|22326737 SEQ IDATP binding/kinase/ IPR011009 Protein kinase- NO: 55 NO: 56 proteinkinase/ like; protein IPR000719 Protein kinase; serine/threonineIPR008271 Serine/threonine kinase/protein- protein kinase, active sitetyrosine kinase IMQ70.5 At5g11860 gi|42573340 SEQ ID gi|42573341 SEQ IDunknown protein IPR004274 NLI interacting NO: 57 NO: 58 factor IMQ70.5At5g11860 gi|30683827 SEQ ID gi|30683828 SEQ ID unknown proteinIPR004274 NLI interacting NO: 59 NO: 60 factor IMQ70.5 At5g11860gi|30683826 SEQ ID gi|15239800 SEQ ID unknown protein IPR004274 NLIinteracting NO: 61 NO: 62 factor IMQ70.6 At5g11870 gi|30683836 SEQ IDgi|15239801 SEQ ID unknown protein NO: 63 NO: 64 IMQ70.7 At5g11880gi|42567795 SEQ ID gi|18416698 SEQ ID diaminopimelate IPR000183Orn/DAP/Arg NO: 65 NO: 66 decarboxylase decarboxylase 2; IPR002986Diaminopimelate decarboxylase; IPR009006 Alanine racemase/group IVdecarboxylase, C-terminal IMQ71.1 At5g12990 gi|18416924 SEQ IDgi|15239964 SEQ ID CLE40 NO: 67 NO: 68 (CLAVATA3/ESR- RELATED 40);receptor binding IMQ71.2 At5g13000 gi|42567812 SEQ ID gi|30684210 SEQ IDATGSL12 (GLUCAN IPR001093 IMP NO: 69 NO: 70 SYNTHASE-LIKEdehydrogenase/GMP 12); 1,3-beta-glucan reductase; synthase/ IPR003440Glycosyl transferase, transferase, family 48; transferring glycosylIPR008207 Hpt groups IMQ71.3 At5g13010 gi|30684214 SEQ ID gi|15239967SEQ ID EMB3011; ATP IPR001410 DEAD/DEAH box NO: 71 NO: 72 binding/ATP-helicase; dependent helicase/ IPR001650 Helicase, C- RNA helicase/terminal; helicase/nucleic acid IPR007502 Helicase- binding associatedregion; IPR011545 DEAD/DEAH box helicase, N-terminal; IPR011709 Proteinof unknown function DUF1605 IMQ72.1 At5g15700 gi|30685486 SEQ IDgi|15242369 SEQ ID DNA binding/DNA- IPR002092 DNA-directed RNA NO: 73NO: 74 directed RNA polymerase, bacteriophage polymerase type IMQ72.2At5g15710 gi|30685493 SEQ ID gi|15242370 SEQ ID unknown proteinIPR001810 Cyclin-like F-box NO: 75 NO: 76 IMQ72.3 At5g15720 gi|18417759SEQ ID gi|18417760 SEQ ID carboxylic ester IPR001087 Lipolytic enzyme,NO: 77 NO: 78 hydrolase/hydrolase, G-D-S-L acting on ester bonds IMQ72.4At5g15725 gi|30685500 SEQ ID gi|18417762 SEQ ID unknown protein NO: 79NO: 80 IMQ72.5 At5g15730 gi|30685503 SEQ ID gi|18417765 SEQ ID ATPbinding/kinase/ IPR011009 Protein kinase- NO: 81 NO: 82 protein kinase/like; protein IPR000719 Protein kinase; serine/threonine IPR008271Serine/threonine kinase/protein- protein kinase, active site tyrosinekinase IMQ73.1 At5g35960 gi|18421515 SEQ ID gi|15239245 SEQ ID ATPbinding/kinase/ IPR011009 Protein kinase- NO: 83 NO: 84 protein kinase/like; protein IPR000719 Protein kinase; serine/threonine IPR008271Serine/threonine kinase/protein- protein kinase, active site tyrosinekinase IMQ73.2 At5g35970 gi|30692867 SEQ ID gi|30692868 SEQ ID ATPbinding/ATP- IPR001093 IMP NO: 85 NO: 86 dependent helicase/dehydrogenase/GMP DNA binding reductase; IPR003593 AAA ATPase; IPR011545DEAD/DEAH box helicase, N-terminal IMQ73.3 At5g35980 gi|42568144 SEQ IDgi|42568145 SEQ ID ATP binding/kinase/ IPR011009 Protein kinase- NO: 87NO: 88 protein kinase/ like; protein IPR000719 Protein kinase;serine/threonine IPR008271 Serine/threonine kinase/protein- proteinkinase, active site tyrosine kinase IMQ73.3 At5g35980 gi|79329053 SEQ IDgi|79329054 SEQ ID ATP binding/kinase/ IPR011009 Protein kinase- NO: 89NO: 90 protein kinase/ like; protein IPR000719 Protein kinase;serine/threonine IPR008271 Serine/threonine kinase/protein- proteinkinase, active site tyrosine kinase IMQ74.1 At5g38530 gi|30693265 SEQ IDgi|15240941 SEQ ID catalytic/pyridoxal IPR001926 Pyridoxal-5′- NO: 91NO: 92 phosphate binding/ phosphate-dependent tryptophan synthaseenzyme, beta subunit; IPR006316 Tryptophan synthase, beta chain-likeIMQ74.2 At5g38540 gi|18421768 SEQ ID gi|15240944 SEQ ID unknown proteinIPR001229 Jacalin-related NO: 93 NO: 94 lectin IMQ74.3 At5g38550gi|42568183 SEQ ID gi|15240945 SEQ ID unknown protein IPR001229Jacalin-related NO: 95 NO: 96 lectin IMQ74.4 At5g38560 gi|30693271 SEQID gi|15240947 SEQ ID ATP binding/kinase/ IPR011009 Protein kinase- NO:97 NO: 98 protein kinase/ like; protein IPR000719 Protein kinase;serine/threonine IPR008271 Serine/threonine kinase/protein- proteinkinase, active site; tyrosine kinase/ IPR003882 Pistil-specificstructural constituent extensin-like protein of cell wall IMQ75.1At5g40300 gi|30693576 SEQ ID gi|15242642 SEQ ID unknown proteinIPR006702 Protein of NO: 99 NO: 100 unknown function DUF588 IMQ75.2At5g40310 gi|18421971 SEQ ID gi|15242645 SEQ ID exonuclease/nucleicIPR006055 Exonuclease; NO: 101 NO: 102 acid binding/zinc ion IPR007087Zinc finger, C2H2- binding type; IPR012337 Polynucleotidyl transferase,Ribonuclease H fold IMQ75.3 At5g40320 gi|18421972 SEQ ID gi|15242647 SEQID protein binding/zinc IPR001965 Zinc finger, PHD- NO: 103 NO: 104 ionbinding type; IPR002219 Protein kinase C, phorbol ester/diacylglycerolbinding; IPR004146 DC1; IPR011424 C1-like IMQ76.1 At5g44180 gi|18422414SEQ ID gi|15241428 SEQ ID transcription factor IPR001356 Homeobox; NO:105 NO: 106 IPR004022 DDT; IPR009057 Homeodomain-like IMQ77.1 At5g62600gi|42568711 SEQ ID gi|42568712 SEQ ID protein transporter IPR001494Importin-beta, N- NO: 107 NO: 108 terminal IMQ77.2 At5g62610 gi|30697712SEQ ID gi|15241896 SEQ ID DNA binding/ IPR001092 Basic helix-loop- NO:109 NO: 110 transcription factor helix dimerisation region bHLH;IPR011598 Helix-loop-helix DNA-binding IMQ78.1 At5g67580 gi|30698319 SEQID gi|30698320 SEQ ID DNA binding/ IPR001005 Myb, DNA-binding; NO: 111NO: 112 transcription factor IPR003216 Linker histone, N- terminal;IPR005818 Histone H1/H5; IPR009057 Homeodomain-like IMQ78.1 At5g67580gi|30698321 SEQ ID gi|15240783 SEQ ID DNA binding/ IPR001005 Myb,DNA-binding; NO: 113 NO: 114 transcription factor IPR003216 Linkerhistone, N- terminal; IPR005818 Histone H1/H5; IPR009057Homeodomain-like IMQ78.2 At5g67590 gi|30698322 SEQ ID gi|15240784 SEQ IDFRO1 IPR006885 ETC complex I NO: 115 NO: 116 (FROSTBITE1) subunitconserved region IMQ78.3 At5g67600 gi|30698323 SEQ ID gi|18425209 SEQ IDrhodopsin-like NO: 117 NO: 118 receptor IMQ78.4 At5g67610 gi|79332806SEQ ID gi|79332807 SEQ ID unknown protein IPR001093 IMP NO: 119 NO: 120dehydrogenase/GMP reductase IMQ78.4 At5g67610 gi|30698324 SEQ IDgi|15240786 SEQ ID unknown protein IPR001093 IMP NO: 121 NO: 122dehydrogenase/GMP reductase

TABLE 3 5. Orthologous Genes: Nucleic Acid/Polypeptide seq. GI# 1. Gene3. Nucleic Acid 4. Polypeptide Nucleic Acid Polypeptide alias 2. Tairseq. GI# seq. GI# GI# GI# Species IMQ66.4 At4g30810 gi|30688809gi|18417667 gi|55773860 gi|55773861 Oryza sativa (japonicacultivar-group) gi|474390 gi|6102957 Hordeum vulgare subsp. vulgaregi|42569653 gi|15227493 Arabidopsis thaliana IMQ67.1 At4g32230gi|18417967 gi|15236713 gi|30689315 gi|30689316 Arabidopsis thalianagi|30689320 gi|22329080 Arabidopsis thaliana gi|79326107 gi|79326108Arabidopsis thaliana gi|533631 gi|1235831 Streptococcus pyogenesgi|687746 gi|687747 Streptococcus pyogenes IMQ67.2 At4g32240 gi|42567326gi|18417969 IMQ67.3 At4g32250 gi|79326107 gi|79326108 gi|30689315gi|30689316 Arabidopsis thaliana gi|30689320 gi|22329080 Arabidopsisthaliana gi|51535562 gi|51535592 Oryza sativa (japonica cultivar-group)gi|22417471 gi|39588025 Caenorhabditis briggsae gi|18420243 gi|18420244Arabidopsis thaliana IMQ67.3 At4g32250 gi|30689315 gi|30689316gi|30689320 gi|22329080 Arabidopsis thaliana gi|51535562 gi|51535592Oryza sativa (japonica cultivar-group) gi|22417471 gi|39588025Caenorhabditis briggsae gi|18420243 gi|18420244 Arabidopsis thalianaIMQ67.3 At4g32250 gi|30689320 gi|22329080 gi|30689315 gi|30689316Arabidopsis thaliana gi|51535562 gi|51535592 Oryza sativa (japonicacultivar-group) gi|22417471 gi|39588025 Caenorhabditis briggsaegi|18420243 gi|18420244 Arabidopsis thaliana IMQ67.4 At4g32260gi|30689323 gi|15236722 gi|394754 gi|394755 Spinacia oleraceagi|29367412 gi|29367413 Oryza sativa (japonica cultivar-group)gi|31980476 gi|31980477 Drosera tokaiensis IMQ68.1 At4g36560 gi|18419861gi|15234443 IMQ68.2 At4g36570 gi|18419863 gi|15234454 gi|42569222gi|15226604 Arabidopsis thaliana gi|30686408 gi|30686409 Arabidopsisthaliana gi|18410812 gi|15222161 Arabidopsis thaliana IMQ68.3 At4g36580gi|18419864 gi|15234455 gi|30680342 gi|18398708 Arabidopsis thalianagi|30686107 gi|22326858 Arabidopsis thaliana gi|50911948 gi|50911949Oryza sativa (japonica cultivar-group) IMQ68.4 At4g36590 gi|18419866gi|15234456 gi|18424355 gi|15239333 Arabidopsis thaliana gi|18378850gi|15223420 Arabidopsis thaliana gi|18408263 gi|15218647 Arabidopsisthaliana IMQ68.5 At4g36600 gi|30690703 gi|30690704 IMQ69.1 At4g38510gi|79326467 gi|79326468 gi|79326459 gi|79326460 Arabidopsis thalianagi|42573220 gi|42573221 Arabidopsis thaliana gi|30691994 gi|15233891Arabidopsis thaliana gi|26986105 gi|26986106 Mesembryanthemumcrystallinum gi|34910487 gi|34910488 Oryza sativa (japonicacultivar-group) gi|459197 gi|459198 Gossypium hirsutum IMQ69.1 At4g38510gi|79326459 gi|79326460 gi|79326467 gi|79326468 Arabidopsis thalianagi|42573220 gi|42573221 Arabidopsis thaliana gi|30691994 gi|15233891Arabidopsis thaliana gi|26986105 gi|26986106 Mesembryanthemumcrystallinum gi|34910487 gi|34910488 Oryza sativa (japonicacultivar-group) gi|459197 gi|459198 Gossypium hirsutum IMQ69.1 At4g38510gi|42573220 gi|42573221 gi|79326467 gi|79326468 Arabidopsis thalianagi|79326459 gi|79326460 Arabidopsis thaliana gi|30691994 gi|15233891Arabidopsis thaliana gi|26986105 gi|26986106 Mesembryanthemumcrystallinum gi|34910487 gi|34910488 Oryza sativa (japonicacultivar-group) gi|459197 gi|459198 Gossypium hirsutum IMQ69.1 At4g38510gi|30691994 gi|15233891 gi|79326467 gi|79326468 Arabidopsis thalianagi|79326459 gi|79326460 Arabidopsis thaliana gi|42573220 gi|42573221Arabidopsis thaliana gi|26986105 gi|26986106 Mesembryanthemumcrystallinum gi|34910487 gi|34910488 Oryza sativa (japonicacultivar-group) gi|459197 gi|459198 Gossypium hirsutum IMQ69.2 At4g38520gi|42573222 gi|42573223 gi|42567510 gi|22329238 Arabidopsis thalianagi|4206121 gi|4206122 Mesembryanthemum crystallinum gi|7768152gi|7768153 Fagus sylvatica gi|30698190 gi|15239244 Arabidopsis thalianaIMQ69.2 At4g38520 gi|42567510 gi|22329238 gi|42573222 gi|42573223Arabidopsis thaliana gi|4206121 gi|4206122 Mesembryanthemum crystallinumgi|7768152 gi|7768153 Fagus sylvatica gi|30698190 gi|15239244Arabidopsis thaliana IMQ69.2 At4g38520 gi|79313268 gi|79313269gi|30684366 gi|18401370 Arabidopsis thaliana gi|50919604 gi|50919605Oryza sativa (japonica cultivar-group) gi|42567510 gi|22329238Arabidopsis thaliana gi|42573222 gi|42573223 Arabidopsis thalianaIMQ69.3 At4g38530 gi|79499921 gi|79499922 gi|30691998 gi|15233918Arabidopsis thaliana gi|18424131 gi|18424132 Arabidopsis thalianagi|1399304 gi|1399305 Glycine max IMQ69.4 At4g38540 gi|30692001gi|15233923 gi|30680885 gi|15239070 Arabidopsis thaliana gi|24745926gi|24745927 Solanum tuberosum gi|30686498 gi|18403916 Arabidopsisthaliana IMQ69.5 At4g38550 gi|42567511 gi|22329240 gi|42570854gi|42570855 Arabidopsis thaliana gi|42570292 gi|42570293 Arabidopsisthaliana gi|42570850 gi|42570851 Arabidopsis thaliana IMQ69.6 At4g38560gi|18420262 gi|15233931 gi|30681216 gi|15226430 Arabidopsis thalianagi|18416479 gi|15238969 Arabidopsis thaliana gi|42567511 gi|22329240Arabidopsis thaliana IMQ69.7 At4g38570 gi|30692008 gi|15233934gi|21745397 gi|21745398 Brassica napus gi|18408916 gi|15220618Arabidopsis thaliana gi|50402823 gi|50402824 Solanum tuberosum IMQ70.1At5g11820 gi|18416683 gi|15239789 gi|18420982 gi|15239570 Arabidopsisthaliana gi|18420983 gi|18420984 Arabidopsis thaliana gi|3097261gi|3097262 Papaver nudicaule IMQ70.2 At5g11830 gi|18416686 gi|15239794gi|18420987 gi|15239591 Arabidopsis thaliana gi|18400929 gi|15233219Arabidopsis thaliana gi|18420988 gi|15239593 Arabidopsis thalianaIMQ70.3 At5g11840 gi|18416688 gi|15239796 gi|50932538 gi|50932539 Oryzasativa (japonica cultivar-group) gi|30698299 gi|15240715 Arabidopsisthaliana gi|3184553 gi|3184557 Synechococcus sp. PCC 7002 IMQ70.4At5g11850 gi|30683823 gi|22326737 gi|50906404 gi|50906405 Oryza sativa(japonica cultivar-group) gi|30698956 gi|15219517 Arabidopsis thalianagi|30685720 gi|22329643 Arabidopsis thaliana IMQ70.5 At5g11860gi|42573340 gi|42573341 gi|30683827 gi|30683828 Arabidopsis thalianagi|30683826 gi|15239800 Arabidopsis thaliana gi|34897435 gi|34897436Oryza sativa (japonica cultivar-group) gi|66803904 gi|66803905Dictyostelium discoideum gi|19612100 gi|55239669 Anopheles gambiae str.PEST IMQ70.5 At5g11860 gi|30683827 gi|30683828 gi|42573340 gi|42573341Arabidopsis thaliana gi|30683826 gi|15239800 Arabidopsis thalianagi|34897435 gi|34897436 Oryza sativa (japonica cultivar-group)gi|66803904 gi|66803905 Dictyostelium discoideum gi|19612100 gi|55239669Anopheles gambiae str. PEST IMQ70.5 At5g11860 gi|30683826 gi|15239800gi|42573340 gi|42573341 Arabidopsis thaliana gi|30683827 gi|30683828Arabidopsis thaliana gi|34897435 gi|34897436 Oryza sativa (japonicacultivar-group) gi|66803904 gi|66803905 Dictyostelium discoideumgi|19612100 gi|55239669 Anopheles gambiae str. PEST IMQ70.6 At5g11870gi|30683836 gi|15239801 gi|18409783 gi|15223970 Arabidopsis thalianagi|14276070 gi|57899179 Oryza sativa (japonica cultivar-group)gi|34905817 gi|34905818 Oryza sativa (japonica cultivar-group) IMQ70.7At5g11880 gi|42567795 gi|18416698 gi|30683117 gi|15231844 Arabidopsisthaliana gi|50907772 gi|50907773 Oryza sativa (japonica cultivar-group)gi|77917618 gi|77920014 Pelobacter carbinolicus DSM 2380 IMQ71.1At5g12990 gi|18416924 gi|15239964 gi|18424867 gi|15238257 Arabidopsisthaliana gi|76791618 gi|76791764 Pseudoalteromonas atlantica T6cgi|40841719 gi|50428681 Oryza sativa (japonica cultivar-group) IMQ71.2At5g13000 gi|42567812 gi|30684210 gi|18390541 gi|18390542 Arabidopsisthaliana gi|30685130 gi|30685131 Arabidopsis thaliana gi|50916184gi|50916185 Oryza sativa (japonica cultivar-group) IMQ71.3 At5g13010gi|30684214 gi|15239967 gi|50937582 gi|50937583 Oryza sativa (japonicacultivar-group) gi|12044831 gi|12044832 Chlamydomonas reinhardtiigi|41053697 gi|41053698 Danio rerio IMQ72.1 At5g15700 gi|30685486gi|15242369 gi|21425638 gi|21425639 Nicotiana sylvestris gi|21425664gi|21425665 Nicotiana tabacum gi|17221594 gi|17221595 Nicotianasylvestris IMQ72.2 At5g15710 gi|30685493 gi|15242370 gi|50929496gi|50929497 Oryza sativa (japonica cultivar-group) gi|42795314gi|42795315 Mimulus lewisii gi|50904332 gi|50904333 Oryza sativa(japonica cultivar-group) IMQ72.3 At5g15720 gi|18417759 gi|18417760gi|30684589 gi|18415211 Arabidopsis thaliana gi|30690834 gi|15220514Arabidopsis thaliana gi|18397193 gi|15220512 Arabidopsis thalianaIMQ72.4 At5g15725 gi|30685500 gi|18417762 gi|30678513 gi|30678514Arabidopsis thaliana IMQ72.5 At5g15730 gi|30685503 gi|18417765 gi|505145gi|505146 Nicotiana tabacum gi|55297390 gi|55297406 Oryza sativa(japonica cultivar-group) gi|50940950 gi|50940951 Oryza sativa (japonicacultivar-group) IMQ73.1 At5g35960 gi|18421515 gi|15239245 gi|30687060gi|30687061 Arabidopsis thaliana gi|22330851 gi|22330852 Arabidopsisthaliana gi|47169781 gi|52075932 Oryza sativa (japonica cultivar-group)IMQ73.2 At5g35970 gi|30692867 gi|30692868 gi|50911900 gi|50911901 Oryzasativa (japonica cultivar-group) gi|50252625 gi|50252651 Oryza sativa(japonica cultivar-group) gi|290918 gi|290919 Mesocricetus auratusIMQ73.3 At5g35980 gi|42568144 gi|42568145 gi|50911864 gi|50911865 Oryzasativa (japonica cultivar-group) gi|50928502 gi|50928503 Oryza sativa(japonica cultivar-group) gi|60467544 gi|60467548 Dictyosteliumdiscoideum IMQ73.3 At5g35980 gi|79329053 gi|79329054 gi|42568144gi|42568145 Arabidopsis thaliana gi|50911864 gi|50911865 Oryza sativa(japonica cultivar-group) gi|50928502 gi|50928503 Oryza sativa (japonicacultivar-group) gi|66810394 gi|66810395 Dictyostelium discoideum IMQ74.1At5g38530 gi|30693265 gi|15240941 gi|51535750 gi|51535758 Oryza sativa(japonica cultivar-group) gi|51535750 gi|51535759 Oryza sativa (japonicacultivar-group) gi|76258946 gi|76258962 Chloroflexus aurantiacus J-10-flIMQ74.2 At5g38540 gi|18421768 gi|15240944 gi|42568183 gi|15240945Arabidopsis thaliana gi|18406629 gi|15218997 Arabidopsis thalianagi|30696152 gi|30696153 Arabidopsis thaliana IMQ74.3 At5g38550gi|42568183 gi|15240945 gi|18421768 gi|15240944 Arabidopsis thalianagi|30696152 gi|30696153 Arabidopsis thaliana gi|18406632 gi|15219000Arabidopsis thaliana IMQ74.4 At5g38560 gi|30693271 gi|15240947gi|30689350 gi|15222649 Arabidopsis thaliana gi|55765765 gi|34894174Oryza sativa (japonica cultivar-group) gi|57900560 gi|57900576 Oryzasativa (japonica cultivar-group) IMQ75.1 At5g40300 gi|30693576gi|15242642 gi|42568716 gi|15241975 Arabidopsis thaliana gi|2598568gi|2598569 Medicago truncatula gi|18404104 gi|15227628 Arabidopsisthaliana IMQ75.2 At5g40310 gi|18421971 gi|15242645 gi|18405598gi|15232874 Arabidopsis thaliana gi|50911998 gi|50911999 Oryza sativa(japonica cultivar-group) gi|34894119 gi|34894120 Oryza sativa (japonicacultivar-group) IMQ75.3 At5g40320 gi|18421972 gi|15242647 gi|30688433gi|22326989 Arabidopsis thaliana gi|18405393 gi|15229491 Arabidopsisthaliana gi|18413571 gi|15234233 Arabidopsis thaliana IMQ76.1 At5g44180gi|18422414 gi|15241428 gi|30690429 gi|15218656 Arabidopsis thalianagi|34911025 gi|34911026 Oryza sativa (japonica cultivar-group)gi|46879193 gi|51854271 Oryza sativa (japonica cultivar-group) IMQ77.1At5g62600 gi|42568711 gi|42568712 gi|62733638 gi|62733653 Lycopersiconesculentum gi|68358861 gi|68358862 Danio rerio gi|32135050 gi|55962517Danio rerio IMQ77.2 At5g62610 gi|30697712 gi|15241896 gi|55419645gi|55419646 Gossypium hirsutum gi|33339702 gi|33339703 Catharanthusroseus gi|42562809 gi|18406408 Arabidopsis thaliana IMQ78.1 At5g67580gi|30698319 gi|30698320 gi|30698321 gi|15240783 Arabidopsis thalianagi|30693241 gi|15229625 Arabidopsis thaliana gi|30694681 gi|18402853Arabidopsis thaliana gi|42571814 gi|42571815 Arabidopsis thalianagi|30694687 gi|30694688 Arabidopsis thaliana gi|34105722 gi|34105723 Zeamays subsp. mays IMQ78.1 At5g67580 gi|30698321 gi|15240783 gi|30698319gi|30698320 Arabidopsis thaliana gi|30693241 gi|15229625 Arabidopsisthaliana gi|30694681 gi|18402853 Arabidopsis thaliana gi|42571814gi|42571815 Arabidopsis thaliana gi|30694687 gi|30694688 Arabidopsisthaliana gi|34105722 gi|34105723 Zea mays subsp. mays IMQ78.2 At5g67590gi|30698322 gi|15240784 gi|50509944 gi|50509945 Oryza sativa (japonicacultivar-group) gi|50938780 gi|50938781 Oryza sativa (japonicacultivar-group) gi|51593212 gi|51593213 Xenopus laevis IMQ78.3 At5g67600gi|30698323 gi|18425209 IMQ78.4 At5g67610 gi|79332806 gi|79332807gi|30698324 gi|15240786 Arabidopsis thaliana gi|42565787 gi|42565788Arabidopsis thaliana gi|41393266 gi|50838976 Oryza sativa (japonicacultivar-group) gi|50917330 gi|50917331 Oryza sativa (japonicacultivar-group) IMQ78.4 At5g67610 gi|30698324 gi|15240786 gi|42565787gi|42565788 Arabidopsis thaliana gi|41393266 gi|50838976 Oryza sativa(japonica cultivar-group) gi|50917330 gi|50917331 Oryza sativa (japonicacultivar-group)

Example 2 Analysis of the Arabidopsis IMQ Sequence

Sequence analyses were performed with BLAST (Altschul et al., 1990, J.Mol. Biol. 215:403-410), PFAM (Bateman et al., 1999, Nucleic Acids Res.27:260-262), INTERPRO (Mulder et al. 2003 Nucleic Acids Res. 31,315-318.), PSORT (Nakai K, and Horton P, 1999, Trends Biochem. Sci.24:34-6), and/or CLUSTAL (Thompson J D et al., 1994, Nucleic Acids Res.22:4673-4680). Conserved domains for each protein are listed in column 8of Table 2.

Example 3

To test whether over-expression of the genes in Tables 1 and 2 alter theseed composition phenotype, protein, digestible protein, oil and fibercontent in seeds from transgenic plants expressing these genes wascompared with protein, digestible protein, oil and fiber content inseeds from non-transgenic control plants. To do this, the genes werecloned into plant transformation vectors behind the strong constitutiveCsVMV promoter and the seed specific PRU promoter. These constructs weretransformed into Arabidopsis plants using the floral dip method. Theplant transformation vector contains a gene, which provides resistanceto a toxic compound, and serves as a selectable marker. Seed from thetransformed plants were plated on agar medium containing the toxiccompound. After 7 days, transgenic plants were identified as healthygreen plants and transplanted to soil. Non-transgenic control plantswere germinated on agar medium, allowed to grow for 7 days and thentransplanted to soil. Transgenic seedlings and non-transgenic controlplants were transplanted to two inch pots that were placed in randompositions in a 10 inch by 20 inch tray. The plants were grown tomaturity, allowed to self-fertilize and set seed. Seed was harvestedfrom each plant and its oil content estimated by Near Infrared (NIR)Spectroscopy using methods previously described. The effect of eachconstruct on seed composition was examined in at least two experiments.

Table 4 lists constructs tested for causing a significant increase inoil, protein, digestible protein or a significant decrease in fiber wereidentified by a two-way Analysis of Variance (ANOVA) test at ap-value≦0.05. The ANOVA p-values for Protein, Oil, Digestible Proteinand Fiber are listed in columns 4-7, respectively. Those with asignificant p-value are listed in bold. The Average values for Protein,Oil, Digestible Protein and Fiber are listed in columns 8-11,respectively and were calculated by averaging the average valuesdetermined for the transgenic plants in each experiment.

TABLE 4 4. 5. 6. ANOVA 7. 10. ANOVA ANOVA Digestible ANOVA Digestible 1.Gene 2. TAIR 3. Construct Protein Oil Protein Fiber 8. Protein 9. OilProtein 11. Fiber IMQ67.1 At4g32230 Pru::At4g32230 0.066 0.175 0.0150.135 102.5% 97.9% 101.4% 98.6% IMQ67.2 At4g32240 CsVMV::At4g32240 0.3680.260 0.355 0.277  98.1% 102.0%   99.3% 99.2% IMQ67.2 At4g32240Pru::At4g32240 0.013 0.027 0.084 0.426 104.2% 95.8% 101.0% 99.3% IMQ67.3At4g32250 CsVMV::At4g32250 0.392 0.134 0.037 0.630 102.1% 97.3% 101.6%99.6% IMQ67.4 At4g32260 Pru::At4g32260 0.024 0.146 0.068 0.181 103.5%96.8% 101.2% 98.5% IMQ69.3 At4g38530 CsVMV::At4g38530 0.115 0.040 0.5070.487 102.2% 97.0% 100.5% 99.3% IMQ69.3 At4g38530 Pru::At4g38530 0.0430.004 0.211 0.114 103.0% 93.7%  98.8% 102.7%  IMQ69.4 At4g38540CsVMV::At4g38540 0.156 0.077 0.127 0.201 102.5% 97.4% 101.2% 99.0%IMQ69.4 At4g38540 Pru::At4g38540 0.015 0.002 0.459 0.044 106.0% 91.2% 99.3% 103.8%  IMQ69.5 At4g38550 CsVMV::At4g38550 0.028 0.003 0.8620.293 102.4% 96.0% 100.1% 101.0%  IMQ71.2 At5g13000 CsVMV::At5g130000.455 0.564 0.006 0.004 100.8% 99.3% 102.6% 96.3% IMQ71.2 At5g13000Pru::At5g13000 0.232 0.155 0.443 0.564 101.7% 98.0% 100.5% 99.5% IMQ71.3At5g13010 CsVMV::At5g13010 0.000 0.002 0.000 0.019 110.8% 91.3% 103.5%98.1% IMQ71.3 At5g13010 Pru::At5g13010 0.025 0.502 0.224 0.617  96.4%101.7%   99.2% 100.6%  IMQ73.3 At5g35980 CsVMV::At5g35980 0.450 0.7210.064 0.007  99.7% 100.3%  102.0% 96.4% IMQ73.3 At5g35980 Pru::At5g359800.461 0.718 0.075 0.017  99.2% 100.7%  101.4% 96.8% IMQ74.1 At5g38530Pru::At5g38530 0.310 0.475 0.002 0.066 101.7% 98.4% 101.9% 98.1% IMQ74.2At5g38540 CsVMV::At5g38540 0.020 0.004 0.801 0.353 103.4% 92.9%  99.7%101.8%  IMQ74.2 At5g38540 Pru::At5g38540 0.956 0.853 0.460 0.514 100.0%99.6% 100.5% 99.3% IMQ75.1 At5g40300 CsVMV::At5g40300 0.025 0.153 0.5370.362  97.6% 102.6%  100.7% 98.8% IMQ75.1 At5g40300 Pru::At5g40300 0.4310.881 0.050 0.044 101.4% 100.1%  101.2% 98.0% IMQ75.2 At5g40310Pru::At5g40310 0.349 0.096 0.015 0.005  99.0% 102.9%  101.9% 97.0%IMQ75.3 At5g40320 CsVMV::At5g40320 1.238 0.106 0.800 0.188 98.55%102.95%  100.20%  98.50%  IMQ75.3 At5g40320 Pru::At5g40320 0.048 0.4440.000 0.000 102.5% 99.0% 103.0% 96.5% IMQ78.4 At5g67610 CsVMV::At5g676100.777 0.586 0.325 0.158 100.4% 101.2%  100.8% 98.1% IMQ78.4 At5g67610Pru::At5g67610 0.009 0.045 0.730 0.026  95.0% 105.2%  100.3% 98.0%

Example 4

To test whether over-expression of the genes identified in Tables 1-4alter the seed composition phenotype, protein, digestible protein, oil,and fiber content in seeds from transgenic plants expressing these genesis compared with protein, digestible protein, oil and fiber content inseeds from non-transgenic control plants. Any one of the genesidentified in Tables 1-4 is used to transform Brassica napus (canola).To do this, the genes are cloned into plant transformation vectorsbehind the strong constitutive CsVMV promoter and the seed specificphaseolin promoter. These constructs (which include a gene encoding aselection agent) are transformed into canola plants.

Transformation of canola is accomplished via Agrobacterium-mediatedtransformation. Seeds are surface-sterilized with 10% commercial bleachfor 10 minutes and rinsed 3 times with sterile distilled water. Theseeds are then placed on one half concentration of MS basal medium(Murashige and Skoog, Physiol. Plant. 15:473-497, 1962) and maintainedunder growth regime set at 25° C., and a photoperiod of 16 hrs light/8hrs dark.

Hypocotyl segments (3-5 mm) are excised from 5-7 day old seedlings andplaced on callus induction medium K1D1 (MS medium with 1 mg/1 kinetinand 1 mg/1 2,4-D) for 3 days as pre-treatment. The segments are thentransferred into a petri plate, treated with Agrobacterium Z7075 orLBA4404 strain containing pDAB721. The Agrobacterium is grown overnightat 28° C. in the dark on a shaker at 150 rpm and subsequentlyre-suspended in the culture medium.

After 30 minute treatment of the hypocotyl segments with Agrobacterium,these are placed back on the callus induction medium for 3 days.Following co-cultivation, the segments are placed on K1D1TC (callusinduction medium containing 250 mg/1 Carbenicillin and 300 mg/1Timentin) for one week of recovery. Alternately, the segments are placeddirectly on selection medium K1D1H1 (above medium with 1 mg/1 selectionagent, for example an herbicide). Carbenicillin and Timentin areantibiotics used to kill the Agrobacterium. The selection agent is usedto allow the growth of the transformed cells.

Callus samples from independent events are tested by PCR. All thesamples tested are positive for the presence of the transformed gene,whereas the non-transformed controls are negative. Callus samples areconfirmed to express the appropriate protein as determined by ELISA.

Callused hypocotyl segments are then placed on B3Z1H1 (MS medium, 3 mg/1benzylamino purine, 1 mg/1 Zeatin, 0.5 μm/1 MES [2-(N-morpholino) ethanesulfonic acid], 5 mg/1 silver nitrate, 1 mg/1 selection agent,Carbenicillin and Timentin) shoot regeneration medium. After shootsstart to regenerate (approximately 3 weeks), hypocotyl segments alongwith the shoots are transferred to B3Z1H3 medium (MS medium, 3 mg/1benzylamino purine, 1 mg/1 Zeatin, 0.5 μm/1 MES [2-(N-morpholino) ethanesulfonic acid], 5 mg/1 silver nitrate, 3 mg/1 selection agent,Carbenicillin and Timentin) for 3 weeks.

Shoots are excised from the hypocotyl segments and transferred to shootelongation medium MESH10 (MS, 0.5 μm/1 MES, 10 mg/1 selection agent,Carbenicillin, Timentin) for 2-4 weeks. The elongated shoots arecultured for root induction on MSI.1 (MS with 0.1 mg/1 Indolebutyricacid). Once the plants have a well-established root system, these aretransplanted into soil. The plants are acclimated under controlledenvironmental conditions in the Conviron for 1-2 weeks before transferto the greenhouse. The transformed T0 plants self-pollinate in thegreenhouse to obtain T1 seed. Transgenic plants are selected at the T1generation based on resistance to a selection agent. T2 seed (from T1plants) is harvested and sown in soil. T2 plants are grown to maturity,allowed to self-fertilize and set seed. T3 seed (from the T2 plants) isharvested in bulk for each line. Seed oil, protein, digestible protein,and fiber values are measured as discussed in Example 1.

We claim:
 1. A transgenic plant, comprising a plant transformationvector comprising a nucleic acid molecule comprising a nucleotidesequence that encodes an IMQ (Improved Meal Quality) polypeptidecomprising the amino acid sequence as set forth in SEQ ID NO: 14, or anIMQ polypeptide comprising an amino acid sequence at least 95% identicalto the amino acid sequence as set forth in SEQ ID NO: 14, whereby thetransgenic plant has an improved meal quality phenotype, relative tocontrol plants.
 2. The transgenic plant of claim 1, wherein the IMQpolypeptide comprises the amino acid sequence as set forth in SEQ ID NO:14.
 3. The transgenic plant of claim 1, which is selected from the groupconsisting of plants of the Brassica species, including canola andrapeseed, soy, corn, sunflower, cotton, cocoa, safflower, oil palm,coconut palm, flax, castor, peanut, wheat, oat, and rice.
 4. Thetransgenic plant of claim 3, wherein the plant is canola.
 5. Thetransgenic plant of claim 1, wherein an improved meal quality phenotypecomprises an increase in available metabolizable energy in meal producedfrom seeds of the transgenic plant, relative to control plants.
 6. Thetransgenic plant of claim 5, wherein an increase in availablemetabolizable energy comprises an altered protein and/or fiber contentin the seeds of the transgenic plant.
 7. The transgenic plant of claim6, wherein the protein content is increased and/or the fiber content isdecreased.
 8. The transgenic plant of claim 5, wherein an increase inavailable metabolizable energy comprises an increased protein content inthe seeds of the transgenic plant.
 9. A plant part obtained from theplant according to claim 1, wherein the plant part comprises saidnucleic acid molecule.
 10. The plant part of claim 9, which is a seed.11. Meal, feed, or food produced from the seed of claim 10, whereby themeal, feed or food from the transgenic plant has improved meal quality,relative to meal, feed or food from a control plant that does notcomprise the nucleic acid molecule.
 12. A method of producing meal,comprising growing the transgenic plant of claim 1, and recovering mealfrom the plant, thereby producing meal.
 13. The method of claim 12,wherein the meal is produced from seeds of the plant.
 14. A method ofproducing an improved meal quality phenotype in a plant, said methodcomprising: a) introducing into progenitor cells of the plant a planttransformation vector comprising a nucleic acid molecule comprising anucleotide sequence that encodes an IMQ polypeptide comprising the aminoacid sequence set forth in SEQ ID NO: 14, or an IMQ polypeptidecomprising an amino acid sequence at least 95% identical to the aminoacid sequence as set forth in SEQ ID NO: 14, and b) growing thetransformed progenitor cells to produce a transgenic plant, wherein thenucleic acid molecule is expressed, and the transgenic plant exhibits animproved meal quality phenotype relative to control plants, therebyproducing the improved meal quality phenotype in the plant.
 15. Themethod of claim 14, wherein the IMQ polypeptide comprises the amino acidsequence as set forth in SEQ ID NO:
 14. 16. A plant obtained by a methodof claim
 14. 17. The plant of claim 16, which is selected from the groupconsisting of plants of the Brassica species, including canola andrapeseed, soy, corn, sunflower, cotton, cocoa, safflower, oil palm,coconut palm, flax, castor, peanut, wheat, oat, and rice.
 18. The plantof claim 17, wherein the plant is canola.
 19. The plant of claim 16,wherein the plant is selected from the group consisting of a plant grownfrom said progenitor cells, a plant that is the direct progeny of aplant grown from said progenitor cells, and a plant that is the indirectprogeny of a plant grown from said progenitor cells.
 20. A feed, meal,grain, food, or seed from the transgenic plant of claim 1, wherein thefeed, meal, grain, food, or seed comprises a nucleic acid moleculecomprising a nucleotide sequence encoding a polypeptide comprising anamino acid sequence at least 95% identical to the amino acid sequence asset forth in SEQ ID NO: 14, whereby the feed, meal, grain, food, or seedfrom the transgenic plant has an improved meal quality phenotype,relative to a control plant that does not comprise the nucleic acid. 21.The feed, meal, grain, food, or seed of claim 20, wherein thepolypeptide is encoded by the nucleotide sequence as set forth in SEQ IDNO:
 13. 22. A feed, meal, grain, food, or seed from the transgenic plantof claim 1, wherein the feed, meal, grain, food, or seed comprises apolypeptide comprising the amino acid sequence as set forth in SEQ IDNO: 14, or a polypeptide comprising an amino acid sequence at least 95%identical to the amino acid sequence as set forth in SEQ ID NO: 14,whereby the feed, meal, grain, food, or seed from the transgenic planthas an improved meal quality phenotype, relative to a control plant. 23.The feed, meal, grain, food, or seed of claim 22, wherein thepolypeptide comprises the amino acid sequence as set forth in SEQ ID NO:14.